Nikon Eclipse Ni-E Instruction Manual
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Nikon Eclipse Ni-E Instruction Manual

Upright motorized microscope
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M571 E
11.8.NF.1 (1/2)
*M571EN01*
Upright Motorized Microscope
Instruction Manual
Operation

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Summary of Contents for Nikon Eclipse Ni-E

  • Page 1 M571 E 11.8.NF.1 (1/2) *M571EN01* Upright Motorized Microscope Instruction Manual Operation...

  • Page 3: Introduction

    Introduction Thank you for purchasing a Nikon product. This instruction manual is written for users of the Nikon ECLIPSE Ni-E microscope. To ensure correct usage, read this manual carefully before operating this product. ● No part of this manual may be reproduced or transmitted in any form without prior written permission from Nikon.

  • Page 4: Contents Of The Manual

    Introduction Contents of the Manual The instruction manual for ECLIPSE Ni-E is provided in two volumes.  Operation (This manual)  Assembly/Maintenance Safety Precautions Assembly Components Troubleshooting Microscopy Operations Maintenance and Storage Before Microscopy Specifications Operation Flowchart Before reading the “Assembly/Maintenance” manual, Bright/Dark-field Microscopy Procedure read the “Safety Precautions”...

  • Page 5: Summary Of Contents

    Summary of Contents Summary of Contents (See the next page for the detailed contents.) Safety Precautions Introduction Notes on Handling the Product Contents of the Manual Symbols Used in This Manual Components (System Configuration and Controls for Each Microscopy) Before Microscopy (Information Setting for Motorized Operation) Operation Flowchart Bright/Dark-field Microscopy...

  • Page 6: Table Of Contents

    Contents Contents Introduction ......................... i Contents of the Manual ....................ii Symbols Used in This Manual ..................ii Summary of Contents ....................... iii Safety Precautions......................vi WARNING and CAUTION Symbols ................vi Meaning of Symbols Used on the Product ..............vi WARNING ....................... vii CAUTION .........................ix Notes on Handling the Product..................
  • Page 7 Contents Operating the Motorized XY Stage..............98 Adjusting the Diopter....................99 Focusing and Centering the Condenser ...............100 Adjusting the Aperture Diaphragm ................101 Using the Condenser ....................103 Compatibility of Condensers with the Magnification of Objectives ....103 Using the NI-CUD-E Motorized Universal Condenser (Dry) ......104 Using the NI-CUD Universal Condenser (Dry)..........

  • Page 8: Safety Precautions

    Safety Precautions Safety Precautions To ensure correct and safe operation, read this manual before using this product. WARNING and CAUTION Symbols Although this product is designed and manufactured to be completely safe during use, incorrect usage or failure to follow the safety instructions provided may cause personal injury or property damage.
  • Page 9 ● To avoid the risk of fire, do not place fabric, must be observed. paper or highly flammable volatile materials Safety is a top design priority for Nikon products. such as gasoline, petroleum benzine, paint Safety is ensured as long as the user observes all...
  • Page 10 Safety Precautions WARNING Hazards of mercury lamps Hazardous sample handling (when using the NI-FLEI Epi-fluorescence This product is intended primarily for microscopic attachment) observations and image capture of cells and The light source used with the epi-fluorescence tissue set on glass slides. attachment (HG Precentered Fiber Illuminator) Check to determine whether a sample is requires special care during handling because of...
  • Page 11 (press to the “O” the product, and contact your nearest Nikon position) and unplugging the power cord from representative. the wall outlet.
  • Page 12 Safety Precautions CAUTION Cautions on sustained observations To relieve fatigue resulting from long observation sessions, limit continuous observations to one hour. Take at least 10 to 15 minutes breaks between observation sessions. Adjust the layout of other equipment used and the height of your chair.

  • Page 13: Notes On Handling The Product

    Notes on Handling the Product Notes on Handling the Product Handling the product carefully ● To avoid splashes, do not use this product near water. This product is a precision instrument. Avoid ● Make sure the ambient temperature is 0 to subjecting it to sudden impacts and shocks.
  • Page 14 Unpacking and unlocking 1) Check the package contents. ● ECLIPSE Ni-E Main Body X1 ● Hex driver X3 Hex wrench X2 ● Toolbox X1 ● Accessory sticker X2 types...

  • Page 15: Components

    This chapter shows figures of devices required for each motorized microscopy with name of components and controls. The figures are examples of basic motorized systems. Manually-operated devices are also available. For the list of components used for the ECLIPSE Ni-E microscope, see Chapter 1 “2 Components List” in the separately provided “Assembly/Maintenance” instruction manual.

  • Page 16: System Configuration And Controls For Bright/Dark-Field Microscopy

    Chapter 1 Components System Configuration and Controls for Bright/Dark-field Microscopy Components Motorized DSC zooming port for quadrocular tube (Model: Camera head NI-RPZ-E) Contact arm (Model: NI-CAM) Motorized quadrocular tilting tube Connector box (Model: NI-TT-E) (Model: NIE-CB) Eyepiece Precentered lamphouse (12V 100W) (Model: NI-LH) Halogen lamp (100W) (Model: PHILIPS 7724 or...

  • Page 17: Controls

    Chapter 1 Components Controls Diopter adjustment ring Condenser focus knob Coarse focus knob Fine focus knob Moving claw of the specimen holder Condenser centering screw NC B OU T Stage Y knob Stage X knob Right control buttons Front control buttons (See “1.3 Buttons”...

  • Page 18: Buttons

    Chapter 1 Components Buttons FUNCTION button (Factory settings: Button 1, 2: Condenser CCW, CW rotation Button 3, 4: Excitation filter wheel CCW, CW rotation Button 5, 6: Barrier filter wheel Display panel CCW, CW rotation) brightness adjustment button Z-RESET button DISPLAY Previous button DISPLAY Next button Switch DISPLAY button...

  • Page 19: Connectors

    ERGO/JOY STAGE (Connect to ergo controller (Connect to motorized XY stage controller) joystick controller) MODEL ECLIPSE Ni-E DIA SHUTTER NIKON CORPORATION TOKYO, JAPAN 9 1 0 0 0 1 M A D E I N J A P A N This device complies with Part 15 of the FCC Rules.

  • Page 20: System Configuration And Controls For Epi-Fluorescence Microscopy

    Chapter 1 Components System Configuration and Controls for Epi-fluorescence Microscopy This section describes the components required for epi-fluorescence microscopy. Fundamental components used in common with bright-field microscopy are omitted. For details, see “1 System Configuration and Controls for Bright/Dark-field Microscopy.” Components Epi-fluorescence attachment (Model: NI-FLEI)
  • Page 21 Chapter 1 Components Controls FL/DIC analyzer slider NI-FA (used for epi-fluorescence/ differential interference contrast microscopy) Field diaphragm lever Aperture diaphragm lever A. ST OP F. ST OP Aperture diaphragm centering screws Field diaphragm centering screws ND filter slider Filter cube name plate window Filter cube replacement slot cover...

  • Page 22: System Configuration And Controls For Differential Interference Contrast Microscopy

    Chapter 1 Components System Configuration and Controls for Differential Interference Contrast Microscopy This section describes the components required for differential interference contrast microscopy. Fundamental components used in common with bright-field microscopy are omitted. For details, see “1 System Configuration and Controls for Bright/Dark-field Microscopy.” Motorized nosepiece (Model: NI-ND6-E) DIC Analyzer slider...

  • Page 23: System Configuration And Controls For Phase Contrast Microscopy

    Chapter 1 Components System Configuration and Controls for Phase Contrast Microscopy This section describes the components required for phase contrast microscopy. Fundamental components used in common with bright-field microscopy are omitted. For details, see “1 System Configuration and Controls for Bright/Dark-field Microscopy.” Ph objectives Motorized universal condenser (Model: NI-CUD-E)
  • Page 24 Chapter 1 Components...

  • Page 25: Microscopy Operations

    Chapter Microscopy Operations This chapter describes the procedure for motorized microscopy operations as shown below. In the examples, the basic system configurations shown in the previous chapter are used. Operation 1: Bright/dark-field microscopy (including the notes on the dark-field microscopy) Operation 2: Epi-fluorescence microscopy (including the notes on the epi-fluorescence/differential interference contrast microscopy and epi-fluorescence/phase contrast microscopy) Operation 3: Differential interference contrast microscopy...

  • Page 26: Before Microscopy

    Chapter 2 Microscopy Operations Before Microscopy Information for motorized operation of the microscope has been set upon shipment. If you start using the microscope with the motorized settings of factory default, turn ON the switch of the Control Box A after the assembly of the microscope to use. However, information on optical elements such as an objective and a filter cube to be attached arbitrarily has “not”...
  • Page 27 Chapter 2 Microscopy Operations Setting Default setting Where to see in this manual (DS-L3 for details) Configured DS-L3 →DS-L3 Chapter 6 “4.1.1 Selecting the Buttons to (See DS-L3 Chapter 2 “3.1 Buttons displayed on the [MICROSCOPE [MICROSCOPE be Displayed” CONTROL] screen CONTROL] Screen (Ni-E)”.) Configured DS-L3...

  • Page 28: Operation Flowchart

    Chapter 2 Microscopy Operations Operation Flowchart The operation flowchart for a microscopy is shown below. See the procedure for each microscopy in the next and subsequent sections for details.
  • Page 29 Chapter 2 Microscopy Operations...
  • Page 30 Chapter 2 Microscopy Operations...
  • Page 31 Chapter 2 Microscopy Operations...

  • Page 32: Bright/Dark-Field Microscopy Procedure

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Bright/Dark-field Microscopy Procedure This section describes the bright-field microscopy procedure which is performed using the microscope's controls and buttons.
  • Page 33 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Display at power ON N i - E V x . x x _ x x x x . x x x x . F 1 When the power is turned on, operation progress is D a t a L o a d i n g .

  • Page 34: Lower The Condenser Slightly From The Uppermost Position

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Adjust the optical path in the tube to direct 100% of the light to the binocular section. Press the BINO optical path button on the front of the main body.

  • Page 35: Fully Open The Field Diaphragm And Aperture Diaphragm

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Fully open the field diaphragm and aperture diaphragm. Press the left side ( mark side) of the DIA field diaphragm and DIA aperture diaphragm button.
  • Page 36 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Set the condenser turret to the [OPEN] (empty: bright-field) position. Press the FUNCTION button 1/2 to turn the condenser turret and display [OPEN] on the front panel of Ni-E.
  • Page 37 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Focus and Optical System Adjustment Bring the 10x objective into the optical path. Press the OBJ CW or CCW button to bring the 10x objective into the optical path.

  • Page 38: Focus On The Sample

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Focus on the sample. Looking through the eyepiece, adjust the brightness for optimum brightness by rotating the dia-illumination brightness control knob.

  • Page 39: Adjust The Interpupillary Distance

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Perform diopter adjustment. Look into the right eyepiece with your right eye and the left eyepiece with your left eye. Turn the diopter adjustment ring of each eyepiece to focus on the specimen.
  • Page 40 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 For dark-field microscopy, set the condenser turret to the [DF] (dark-field module) position (this step is not required for bright-field microscopy). Press the FUNCTION button 1/2 to turn the condenser turret and display [DF] on the front panel of Ni-E.

  • Page 41: Oil Immersion

    Press the OBJ CW or CCW button to bring the desired objective into the optical path. Oil immersion When using an oil immersion type objective, apply Nikon specified non-fluorescence immersion oil between the specimen and the objective. (See Chapter 3 “14 Bringing desired objective into optical path Oil/Water Immersion”.)

  • Page 42: Adjust The Aperture Diaphragm

    O P E N A S 1 2 . 3 m m F S 3 0 . 0 m m resolution, Nikon does not recommend setting the aperture diaphragm to less than 60% of the numerical aperture of the objective.

  • Page 43: Adjust The Field Diaphragm

    Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Adjust the field diaphragm. Press the DIA field diaphragm button to adjust the field diaphragm so that it circumscribes the field of view. Size of the field diaphragm Normally, the field diaphragm should be adjusted so that it circumscribes the field of view.
  • Page 44 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Adjust the brightness. Adjust the brightness of the image using the ND filter. Press the ND filter IN/OUT switch to the [IN] (lower) side to bring the filter into the optical path.
  • Page 45 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Viewing the Specimen View the specimen. To observe another part of the sample, rotate the stage knobs to find the position to observe.
  • Page 46 Chapter 2 Microscopy Operations Focus and Optical Micro- Preparation Bright/Dark-field System Adjustment scopy Microscopy 9 10 11 12 13 14 15 16 17 18 19 Turn off the power. Turn off the power switches (press to the “O” position) on the control box A and connected motorized devices.

  • Page 47: Epi-Fluorescence Microscopy Procedure

    Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Epi-fluorescence Microscopy Procedure This section describes the epi-fluorescence microscopy procedure which is performed using the microscope's controls and buttons. Precautions when switching from epi-fluorescence microscopy to differential interference contrast microscopy or phase contrast microscopy are also described.
  • Page 48 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Close the shutter and block the illumination path. Press the [FL SHUTTER] button and close the shutter in the motorized epi-fluorescence cube turret. The shutter is closed when the shutter button is lit.
  • Page 49 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Bring the filter cube into the optical path. Press the FL CUBE CW/CCW button to bring the desired filter cube into the optical path. Pressing the FL CUBE CW button turns the turret by one address in the clockwise direction (as viewed from above), while the CCW button turns the turret in the...
  • Page 50 OBJ CCW button turns the nosepiece in the counterclockwise direction. Bringing desired objective into optical path Oil immersion When using an oil immersion type objective, apply Nikon 4 0 x _ _ _ _ _ _ _ _ _ 0 .
  • Page 51 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Open the shutter. Press the [FL SHUTTER] button and open the shutter in the motorized epi-fluorescence cube turret which is closed in procedure 2.
  • Page 52 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Focus and Optical System Adjustment Focus on the sample. Focus on the sample by rotating the focus knob. (See Chapter 3 “4.1 Proper Focusing Procedure”.) Z-RESET The Z-axis coordinate value can be reset to zero.
  • Page 53 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Adjust the field diaphragm of the epi-fluorescence attachment. Use the field diaphragm lever of the epi-fluorescence attachment to adjust the field diaphragm so that it circumscribes the field of view.
  • Page 54 For fluorescence observations, press the microscope's dia-illumination ON/OFF switch to “OFF” to make the diascopic image disappear. Bright ambient lights will make it difficult to view the image. Nikon recommends keeping the room dark during fluorescence observations. NI-FA FL/DIC Analyzer Slider/Dummy Slider...
  • Page 55 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Turn off the power. Turn off the power switches (press to the “O” position) on the control box A and connected motorized devices. Power LED OFF (Each power LED turns off.) Power switch...
  • Page 56 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Switch from Epi-Fluorescence Microscopy to Bright-field Microscopy When switching from epi-fluorescence to bright-field microscopy, close the epi-fluorescence illumination path shutter and block the illumination of the mercury lamp.
  • Page 57 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Switch from Differential Interference Contrast Microscopy to Epi-Fluorescence Microscopy Epi-fluorescence and differential interference contrast methods can be used concurrently to cover the shortcomings of each.
  • Page 58 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ When differential interference contrast microscopy is finished, go to epi-fluorescence microscopy. • Press the dia-illumination ON/OFF switch to turn off the lamp and make the diascopic image disappear.
  • Page 59 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Concurrent Use of Epi-Fluorescence/Differential Interference Contrast Microscopy FL/DIC analyzer slider By bringing the following optical elements into the optical path, epi-fluorescence and differential interference contrast can be simultaneously observed by microscope: F.
  • Page 60 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Switch from Phase Contrast Microscopy to Epi-Fluorescence Microscopy Epi-fluorescence and phase contrast methods can be used concurrently to cover the shortcomings of each. For example, when locating the target, phase contrast microscopy can be used instead of epi-fluorescence microscopy which tends to cause decoloration of the specimen.
  • Page 61 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ • Rotate the epi-fluorescence cube turret to bring the desired filter cube into the optical path. Bringing the filter cube into the optical path •...
  • Page 62 Chapter 2 Microscopy Operations Preparation Focusing Microscopy Epi-fluorescence Microscopy Procedure 9 10 11 12 13 14 ■ ■ ■ ■ ■ Concurrent Use of Epi-Fluorescence/Phase Contrast Microscopy By bringing the following optical elements into the optical path, epi-fluorescence and phase contrast can be F.

  • Page 63: Differential Interference Contrast Microscopy Procedure

    Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Differential Interference Contrast Microscopy Procedure This section describes the differential interference contrast microscopy procedure which is performed using the microscope's controls and buttons.
  • Page 64 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Display at power ON N i - E V x .
  • Page 65 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Adjust the optical path in the tube to direct 100% of the light to the binocular section. Press the BINO optical path button on the front of the main body.
  • Page 66 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Fully open the field diaphragm and aperture diaphragm. Press the left side ( mark side) of the DIA field diaphragm and DIA aperture diaphragm button.
  • Page 67 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Set the condenser turret to the [OPEN] (empty: bright-field) position. Press the FUNCTION button 1/2 to turn the condenser turret and display [OPEN] on the front panel of Ni-E.
  • Page 68 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Remove the polarizer, analyzer, lambda plate, and DIC slider from the optical path. Loosen the polarizer rotating clamp screw, turn the upper D - D P JAPAN...
  • Page 69 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Place a specimen on the stage, and move the stage to bring the target into view. Open the claw of the specimen holder's moving part and place the specimen onto the stage, gently stowing the claw back to fix the specimen.
  • Page 70 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Focus on the sample. Looking through the eyepiece, adjust the brightness for optimum brightness by rotating the dia-illumination brightness control knob.
  • Page 71 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Perform diopter adjustment. Look into the right eyepiece with your right eye and the left eyepiece with your left eye.
  • Page 72 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Adjust the orientation (vibration direction) of the polarizer and analyzer. This adjustment is performed to determine the basic performance of differential interference contrast method.
  • Page 73 Press the OBJ CW or CCW button to bring the desired objective into the optical path. Oil immersion When using an oil immersion type objective, apply Nikon specified non-fluorescence immersion oil between the specimen and the objective. (See Chapter 3 “14 Bringing desired DIC objective into optical path Oil/Water Immersion”.)
  • Page 74 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Bring the DIC module (on the condenser) into the optical path. Press the FUNCTION button 1/2 to turn the condenser turret and bring the DIC module appropriate for the objective into the optical path.
  • Page 75 A S 1 2 . 8 m m _ _ _ F S 3 0 . 0 m m diaphragm opening will degrade the image resolution, Nikon does not recommend setting the aperture diaphragm to less than 60% of the Aperture numerical aperture of the objective.
  • Page 76 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Adjust the field diaphragm. Press the DIA field diaphragm button to adjust the field diaphragm so that it circumscribes the field of view.
  • Page 77 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Adjust the brightness. Adjust the brightness of the image using the ND filter. Press the ND filter IN/OUT switch to the [IN] (lower) side to bring the filter into the optical path.
  • Page 78 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Viewing the Specimen View the specimen. To observe another part of the sample, rotate the stage λ...
  • Page 79 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Turn off the power. Turn off the power switches (press to the “O” position) on the control box A and connected motorized devices.
  • Page 80 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Differential Interference System Adjustment Contrast Microscopy 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 ■ ■ Switch from Differential Interference Contrast Microscopy to Bright-field Microscopy Loosen the polarizer rotation clamp screw and rotate the upper part of the rotatable polarizer unit clockwise to remove the polarizer from the optical path.

  • Page 81: Phase Contrast Microscopy Procedure

    Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Phase Contrast Microscopy Procedure This section describes the phase contrast microscopy procedure which is performed using the microscope's controls and buttons.
  • Page 82 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Display at power ON N i - E V x . x x _ x x x x . x x x x . F 1 When the power is turned on, operation progress is D a t a L o a d i n g .
  • Page 83 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Adjust the optical path in the tube to direct 100% of the light to the binocular section. Press the BINO optical path button on the front of the main body.
  • Page 84 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Fully open the field diaphragm and aperture diaphragm. Press the left side ( mark side) of the DIA field diaphragm and DIA aperture diaphragm button.
  • Page 85 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Set the condenser turret to the [OPEN] (empty: bright-field) position. Press the FUNCTION button 1/2 to turn the condenser turret and display [OPEN] on the front panel of Ni-E.
  • Page 86 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Focus and Optical System Adjustment Bring the 10x objective (Ph1) into the optical path. Press the OBJ CW or CCW button to bring 10x objective (Ph1) into the optical path.
  • Page 87 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Focus on the sample. Looking through the eyepiece, adjust the brightness for optimum brightness by rotating the dia-illumination brightness control knob.
  • Page 88 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Perform diopter adjustment. Look into the right eyepiece with your right eye and the left eyepiece with your left eye.
  • Page 89 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Bring the PH module (Ph1) into the optical path. Press the FUNCTION button 1/2 to rotate the condenser turret to display [Ph1] on the display panel at the front side of the Ni-E main body.
  • Page 90 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Center the PH module (Ph annular diaphragm). To optimize the phase contrast effect, it is important to Centering telescope Eyepiece properly overlap the phase plate of the objective with the...
  • Page 91 Press the OBJ CW or CCW button to bring the desired Ph objective into the optical path. Oil immersion When using an oil immersion type objective, apply Nikon specified non-fluorescence immersion oil between the specimen and the objective. (See Chapter 3 “14 Bringing desired Ph objective into optical path Oil/Water Immersion”.)
  • Page 92 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Adjust the PH module in the condenser with the Ph objective to be used. Press the FUNCTION button 1/2 to turn the condenser turret and bring the PH module with the same Ph code as the objective into optical path.
  • Page 93 A S 1 2 . 3 m m _ _ _ F S 3 0 . 0 m m diaphragm opening will degrade the image resolution, Nikon does not recommend setting the aperture diaphragm to less than 60% of the Aperture numerical aperture of the objective.
  • Page 94 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Adjust the field diaphragm. Press the DIA field diaphragm button to adjust the field diaphragm so that it circumscribes the field of view.
  • Page 95 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Adjust the brightness. Adjust the brightness of the image using the ND filter. Press the ND filter IN/OUT switch to the [IN] (lower) side to bring the filter into the optical path.
  • Page 96 Chapter 2 Microscopy Operations Focus and Optical Preparation Microscopy Phase Contrast System Adjustment Microscopy Procedure 9 10 11 12 13 14 15 16 17 18 19 20 21 22 ■ ■ Turn off the power. Turn off the power switches (press to the “O” position) on the control box A and connected motorized devices.

  • Page 97: Individual Operations

    Chapter Individual Operations This chapter describes how to use the display panel, operation buttons and each part/device of the Ni-E main body in detail.

  • Page 98: Display Panel Details

    Chapter 3 Individual Operations Display Panel Details The 2-line, 24-character fluorescent display panel on the front of the main body can display the following information: Power ON display When the power is turned on, operation progress is N i - E V x .
  • Page 99 Chapter 3 Individual Operations Microscope State Display Pattern 1 Objective, Z-axis position (unit: um) _ _ _ _ _ _ _ _ _ 0 . 0 0 0 u m Bottom Dia-illumination status, field diaphragm status (unit: D I A : O N _ _ _ _ _ _ _ _ _ _ F S 3 0 .
  • Page 100 Chapter 3 Individual Operations Contents of the “microscope state” display are as follows. Microscope State Display Contents ]： Blank: Achromat Objective 0 0 0 x ]： Apo TIRF A T 0 0 0 x ]： Plan 0 0 0 x ]：...

  • Page 101: Using Operation Buttons On Ni-E

    Chapter 3 Individual Operations Using Operation Buttons on Ni-E Various operation buttons are provided on the front, right, and left of the Ni-E main body. The following describes the function of each button (factory default setting): Front Operation Button Front Operation Button Name Function Display panel...

  • Page 102: Right Operation Button

    Chapter 3 Individual Operations Right Operation Button Right Operation Button Name Function FL CUBE CW button* Rotate the 1st layer motorized epi-fluorescence cube turret by one address clockwise (as viewed from above). FL CUBE CCW button* Rotate the 1st layer motorized epi-fluorescence cube turret by one address counterclockwise (as viewed from above).

  • Page 103: Left Operation Button

    Chapter 3 Individual Operations Left Operation Button Left Operation Button Name Function OBJ CW button* Rotate the motorized nosepiece by one address clockwise (as viewed from above). OBJ CCW button* Rotate the motorized nosepiece by one address counterclockwise (as viewed from above).

  • Page 104: Adjusting The Brightness Of A Diascopic Image

    Chapter 3 Individual Operations Adjusting the Brightness of a Diascopic Image The brightness of a diascopic image can be adjusted by changing the lamp voltage or by using ND filters. Adjustment by Lamp Voltage Turning the dia-illumination brightness control knob changes the voltage of the lamp to change the brightness of the diascopic image.

  • Page 105: Adjustment With Nd Filters

    Chapter 3 Individual Operations Adjustment with ND Filters There are two methods for adjusting the brightness with ND filters: bringing/removing the main body's internal ND filters in/from the optical path, or operating the motorized ND filter wheel attached between the main body and dia-illumination. Using ND filters in the main body ND filters are used to adjust light intensity.

  • Page 106: Focusing On The Sample (Vertical Stage Movement)

    Chapter 3 Individual Operations Focusing on the Sample (Vertical Stage Movement) Focus adjustment is performed using the focus knobs on the Ni-E main body, on the ergo controller or on the joystick controller. The following instructions assume the use of the focus knobs on the main body. For details on using the ergo controller or joystick controller, see “20 Using the Ergo Controller”...

  • Page 107: Focus Knobs On The Main Body

    Chapter 3 Individual Operations Enabling/disabling the operation of the elevating section You can disable the operation of the elevating section. As factory default setting, a focus knob is used to drive the elevating section, but you can “disable” the setting when an application software such as NIS-E is used to control the elevating section or you wan to prevent unintended behavior of the elevating section, including an accidental contact of the focus knob of Ni-E microscope or ergo controller (or joystick).

  • Page 108: Resetting The Z-Axis Coordinate

    Chapter 3 Individual Operations Position Exchange of the Fine Focus Knob The fine focus knobs, located on both sides of the main body, are flat on one end, and convex on the other. The fine focus knob on each side is attached with a magnet, and can easily be swapped.

  • Page 109: Refocusing

    Chapter 3 Individual Operations Refocusing The stage can be retracted to a preset retracting position, and then restored back to the original position. Pressing the Escape button lowers the stage to the preset retracting position while saving the position where the Escape button is pressed.

  • Page 110: Bringing The Target Into The Optical Path (Horizontal Stage Movement, Rotation)

    Chapter 3 Individual Operations Bringing the Target into the Optical Path (Horizontal Stage Movement, Rotation) When using a manual rotatable ceramic coated stage, use the stage X, Y knobs or rotate the top stage plate to move the specimen and bring the target into the optical path. (So that the sample sealed under the cover glass will be lighted.) When using a motorized XY stage, use the ergo controller or joystick to control the horizontal stage movement.
  • Page 111 Chapter 3 Individual Operations Stage rotation The rotatable ceramic coated stage allows you to rotate the specimen under observation. To adjust the orientation of the specimen, loosen the rotation clamp screw on the stage, and then rotate the stage as necessary. This capability is useful in aligning the specimen with the shearing direction to enhance 195°...

  • Page 112: Operating The Motorized Xy Stage

    Chapter 3 Individual Operations Adjusting the knob rotation torque When the X knob and Y knob are moved to the top and bottom positions, the torque adjustment screws can be found between the knobs. Turning the torque adjustment screw to move it closer to the respective knob increases rotational torque.

  • Page 113: Adjusting The Diopter

    Chapter 3 Individual Operations Adjusting the Diopter The diopter adjustment ring on an eyepiece can be adjusted to match the eyesight of your right and left eyes. A properly adjusted diopter compensates for differences in visual acuity between the right and left eyes of a person, making binocular observation easier.

  • Page 114: Focusing And Centering The Condenser

    Chapter 3 Individual Operations Focusing and Centering the Condenser Adjust the condenser position so that the light passing through the condenser forms an image at the correct position (center of the optical path) on the surface of the sample. Follow Steps 1 through 11 in Chapter 2, 3 “Bright/Dark-field Microscopy Procedure”...

  • Page 115: Adjusting The Aperture Diaphragm

    70～80 images with suitable contrast. Since an excessively small aperture diaphragm opening will degrade image resolution, Nikon does not recommend setting the aperture diaphragm to less than 60% of the numerical aperture of the objective. Proper size of the aperture diaphragm A small aperture diaphragm opening reduces resolution and brightness but increases contrast and depth of focus.
  • Page 116 Or, pull out the eyepiece and look into the tube to view the pupil Plan 40X of the objective (bright area when the aperture diaphragm is fully 40x / 0.75 Nikon JAPAN opened). Adjust the aperture diaphragm lever so that the size of Plan 40x 40x / 0.75...

  • Page 117: Using The Condenser

    Chapter 3 Individual Operations Using the Condenser The Ni-E basically uses a motorized universal condenser that supports all microscopy methods (bright-field, dark-field, differential interference contrast, and phase contrast). It can also use various manually-operated condensers such as manual universal condenser. Use the condenser appropriate for the microscopy method and objective.

  • Page 118: Using The Ni-Cud-E Motorized Universal Condenser (Dry)

    Chapter 3 Individual Operations Using the NI-CUD-E Motorized Universal Condenser (Dry) NI-CUD-E is a dry type motorized universal condenser that supports various microscopy methods. ■ Optical module The turret of the universal condenser has one empty hole and six holes for attaching optical modules. The hole at address 1 is empty and no optical module can be attached here.

  • Page 119: Using The Ni-Cud Universal Condenser (Dry)

    Chapter 3 Individual Operations Aperture diaphragm at phase contrast microscopy When using the NI-CUD-E for phase contrast microscopy, fully open the aperture diaphragm. Stopping down the aperture diaphragm may block the PH module's annular diaphragm, disabling phase contrast microscopy. ■ Condenser focus knob On the motorized universal condenser, the condenser focusing unit is integrated with the condenser and a condenser focus knob is attached on the left and right sides of the focusing unit.
  • Page 120 Chapter 3 Individual Operations  PH module (required for phase contrast microscopy) PH module includes [D-C PH-1], [D-C PH-2], and [D-C PH-3]. Select the PH module with the same PH code as the one indicated on the PH objective.  Dark-field module (required for dark-field microscopy) The dark-field module [D-C DF] can be used with any objective with NA (numerical aperture) of 0.7 or less.

  • Page 121: Using The D-Cuo Dic Condenser (Oil)

    Chapter 3 Individual Operations Using the D-CUO DIC Condenser (Oil) The D-CUO is an oil type condenser that provides best results for differential interference contrast microscopy with oil Aperture diaphragm lever immersion objectives. When using it, always apply immersion oil between the slide glass and the tip of the condenser lens.

  • Page 122: Adjusting The Field Diaphragm

    Chapter 3 Individual Operations Adjusting the Field Diaphragm The field diaphragm is used to restrict illumination to the area of the sample being viewed. Operate the DIA field diaphragm button to change the size of the field diaphragm. Press the left side ( mark side) of the DIA field diaphragm button to open, and the right side ( mark side) to close the...

  • Page 123: Switching The Objective

    Chapter 3 Individual Operations Switching the Objective 11.1 Motorized Nosepiece Operation To switch objectives attached to a motorized nosepiece, press the OBJ CW/CCW button. The nosepiece rotates to the adjacent address clockwise/counterclockwise each time the OBJ CW/CCW button is pressed. Attachment information display for the objective is disabled by factory default.
  • Page 124 Chapter 3 Individual Operations In order to set the rotation stop, a motorized nosepiece must be attached. “Rotation stop” is disabled by factory default. Configuration is performed on DS-L3. (See “23 Operation on DS-L3” - “23.1 Setting Up the Microscope - (4-3) Z Position Nosepiece Rotation Stop”...

  • Page 125: Intelligent/Manual Nosepiece Operation

    Chapter 3 Individual Operations Various interlocking settings along with the switch of objectives The following can be interlocked when switching objectives:  Retraction of the elevating section (stage)  Adjustment of the size of the aperture diaphragm  Adjustment of the size of the field diaphragm ...

  • Page 126: Switching The Optical Path Of The Tube

    Chapter 3 Individual Operations Switching the Optical Path of the Tube 12.1 Optical Path Switching of the Motorized Quadrocular Tilting Tube The motorized quadrocular tilting tube has three optical paths, directed to the binocular, tube adapter, and rear port. Pressing Tube an optical path button directs 100% of the light to the corresponding optical path.

  • Page 127: Switching The Optical Path In A Manually-Operated Tube

    Chapter 3 Individual Operations Switching the Optical Path in a Manually-Operated Tube 12.2 On a tube with an optical path switching lever, distribution of light can be controlled by operating the lever (or the dial, in the case of a quadrocular tilting tube). Optical path Optical path switching dial...

  • Page 128: Adjusting The Binocular Section

    Chapter 3 Individual Operations Adjusting the Binocular Section 13.1 Adjusting with the Binocular Section of the Tube The binocular section of an ergonomic tube or a quadrocular tilting tube can be tilted. With the ergonomic tube, the binocular section can be extended along the optical axis. Adjust the position of the binocular section for most comfortable viewing.

  • Page 129: Oil/Water Immersion

    After use, thoroughly wipe off all oil, and make sure that no oil remains on the tips of other objectives. Additionally, carefully wipe off the oil from the condenser. Use petroleum benzine to wipe off the immersion oil. For optimum results, Nikon recommends cleaning with absolute alcohol (ethyl or methyl alcohol) after cleaning with petroleum benzine.
  • Page 130 Chapter 3 Individual Operations Water immersion An objective with a "WI" or "W" marking is a water-immersion objective that can focus on the sample at an operating distance under water. This objective is used by filling the chamber with water (distilled water or physiological saline) and dipping the tip of the objective into the water.

  • Page 131: Dark-Field Microscopy

    Chapter 3 Individual Operations Dark-field Microscopy While the dark-field microscopy procedures provided in Chapter 2 uses a motorized universal condenser, dark-field microscopy can also be conducted using a dark-field condenser (dry or oil). Note the following when using these condensers for dark-field microscopy. Using a dark-field condenser (dry or oil) Focus and center a dark-field condenser as described below.

  • Page 132: Epi-Fluorescence Microscopy

    Chapter 3 Individual Operations Epi-fluorescence Microscopy Epi-fluorescence microscopy used for the observation of a fluorescence image requires a brighter illuminator including an optical element such as a filter or mercury lamp. In Ni-E, an epi-fluorescence cube turret (motorized, intelligent, or manual) to which a filter cube is attached, HG precentered fiber illuminator (motorized or manual) which uses mercury lamp as the illuminator, and epi-fluorescence attachment equipped with an ND filter, aperture diaphragm, and field diaphragm are used.
  • Page 133 Chapter 3 Individual Operations Using double layered epi-fluorescence cube turrets Two epi-fluorescence cube turrets can be used by layering them on top of each other. Attach an attachment using a laser as an illuminator on the 1st layer, which should be used for light stimulation when using a double-decked turret.

  • Page 134: Selecting Filters

    Chapter 3 Individual Operations Selecting Filters 16.2 A filter cube consists of three types of optical components: an excitation filter (EX filter), a barrier filter (BA filter), and a dichroic mirror (DM). Select the filter cube with the desired combination of optical components to suit the characteristics of the sample Barrier filter U V - 2 A...
  • Page 135 – for example, TRITC in the case of FITC and TRITC. This will result in very dark TRITC fluorescent images. For such cases, Nikon recommends using Wavelength multiband filters. Long-pass filter (Both FITC and TRITC images are visible.)

  • Page 136: Protecting The Sample And Preventing It From Decoloration (Using The Shutter)

    Chapter 3 Individual Operations Protecting the Sample and Preventing It from Decoloration (Using the Shutter) 16.3 If the sample is continuously exposed to the strong light of the mercury lamp used for the epi-fluorescence attachment, it may become damaged or decolorized. Be sure to close the shutter when suspending microscopy or when pausing epi-fluorescence microscopy to perform microscopy with diascopic light.
  • Page 137 Chapter 3 Individual Operations ■ Motorized shutter For Ni-E, the motorized shutter can be attached to the epi-fluorescence side or dia-illumination side to provide a fast opening/closing of an epi-fluorescence illumination path or EPI motorized shutter dia-illumination path. On the epi-fluorescence side, attach the shutter between the epi-fluorescence attachment and the HG adapter of the HG precentered fiber illuminator.

  • Page 138: Adjusting The Brightness Of The Fluorescent Image (Using Nd Filters And The Aperture Diaphragm)

    Chapter 3 Individual Operations Manually opening and closing the shutter of a unit Use a shutter open/close lever on the lower front of the epi-fluorescence cube turret to open/close the shutter built in a manual/intelligent epi-fluorescence cube turret. Lever position [C]: Shutter closed.
  • Page 139 Chapter 3 Individual Operations ND filters in the motorized HG precentered fiber illuminator The ND value of the ND filters in the motorized HG precentered fiber illuminator can be controlled to adjust the brightness incrementally. Z Speed XY Speed Brightness can be adjusted in six steps, with the relative light FL cube OBJ intensity cycling in the following order each time the switch is pressed: 1 →...

  • Page 140: Restricting The Illumination To The Area Of The Specimen Being Viewed (Centering And Adjusting The Field Diaphragm)

    Chapter 3 Individual Operations Restricting the Illumination to the Area of the Specimen being Viewed 16.5 (Centering and Adjusting the Field Diaphragm) The field diaphragm is used to restrict illumination to the area of the sample being viewed. Turning the field diaphragm lever of the epi-fluorescence attachment changes the size of the field diaphragm.

  • Page 141: Changing The Waveform Characteristics Of The Excitation Light (D-Fb Excitation Balancer)

    Chapter 3 Individual Operations Changing the Waveform Characteristics of the Excitation Light (D-FB Excitation Balancer) 16.6 By attaching the D-FB Excitation Balancer to the epi-fluorescence attachment, the waveform characteristics of the excitation light can be changed arbitrarily. The balancer is to be used together with a dual-band filter cube.

  • Page 142: Switching The Excitation Filter (Motorized Excitation Filter Wheel)

    Chapter 3 Individual Operations ■ Objectives The excitation balancer should be used in combination with the following objectives. Use of other objectives may result in unevenness in the field of view. Plan fluor 40x/0.75 40xH/1.3 100xH/1.3 S fluor 40x/0.9 40xH/1.3 100xH/1.3 Plan apochromat 40x/0.95...

  • Page 143: Switching The Barrier Filter (Motorized Barrier Filter Wheel)

    Chapter 3 Individual Operations Switching the Barrier Filter (Motorized Barrier Filter Wheel) 16.8 By attaching a motorized barrier filter wheel between the epi-fluorescence cube turret and the quadrocular tilting tube, switching of barrier filters can be motorized. The wheel has one empty hole and seven slots.

  • Page 144: Differential Interference Contrast Microscopy

    Chapter 3 Individual Operations Differential Interference Contrast Microscopy The differential interference contrast microscopy in which a transparent or achromatic sample is observed with higher contrast requires an analyzer, polarizer, and DIC slider on the objective and DIC module on the condenser side. The analyzer and DIC slider for the objective are inserted to slots of the nosepiece when used, therefore use a DIC nosepiece with slots for the differential interference contrast microscopy.

  • Page 145: Using Optical Elements

    Chapter 3 Individual Operations Using Optical Elements 17.2 ■ DIC module for the condenser There are various kinds of DIC modules: [D-C DIC N1 DRY], [D-C DIC N2 DRY] and [D-C DIC NR DRY] to be attached to the universal condenser, and [D-C DIC N2 OIL] and [D-C DIC NR OIL] to be attached to the DIC condenser. Note that an inappropriate combination may result in an inability to produce an interference image, or a significant degradation of contrast.

  • Page 146: Phase Contrast Microscopy

    Chapter 3 Individual Operations Phase Contrast Microscopy Phase contrast microscopy is suitable for observation of clear and colorless specimens, undyed or lightly colored specimens, decolored specimens, and ultrathin slices for electron microscopes. The phase contrast method is not suitable for graded or hard dyed specimens. Optical elements necessary for the microscopy is a Ph objective (phase plate) and a PH module (Ph annular diaphragm) on the condenser.

  • Page 147: Using Optical Elements

    Chapter 3 Individual Operations There is a Ph objective used for APC (Apodized Phase Contrast) microscopy that reduces halos with enhanced contrast of a minute structure compared to the above phase contrast microscopy. See the table below for the use properties of Ph objectives for APC observation.

  • Page 148: Useful Functions

    Chapter 3 Individual Operations Useful Functions 19.1 MODE “MODE” saves and loads a “microscopy state” (position and status of motorized devices) and is useful for an observation when switching between different microscopy state. To use this function, you must have registered a set of motorized units for loading in each mode. More than one motorized unit can be registered for one mode with up to 8 modes available for setting.
  • Page 149 Chapter 3 Individual Operations Now MODE1 has been registered and the registered mode name is displayed on the [MODE1] button. [SAVE MODE1] button is shown on the right of the mode name button. Repeat the same procedure for MODE2 registration. (4) MODE1 and MODE2 have been registered Saving a mode Save the mode on the [MICROSCOPE SETUP] screen.
  • Page 150 Chapter 3 Individual Operations Loading a mode Press [FUNCTION1] and [FUNCTION2] buttons alternately. This assumes that a load function for MODE 1 and MODE 2 is allocated to FUNCTION 1 button and FUNCTION 2 button respectively. The MODE load function is not set to a FUNCTION button in Ni-E Loading MODE as default.
  • Page 151 Chapter 3 Individual Operations Motorized Unit that Save/Loads the State with MODE Function Setting items Stored Information CONDEN. Condenser module of the motorized universal condenser in the optical path FL TURRET Filter cube of the motorized epi-fluorescence cube turret in the optical path FL [SHUTTER] Opening/closing of the motorized epi-fluorescence cube turret built-in shutter FL 2nd...

  • Page 152: Interlocking Function

    Chapter 3 Individual Operations Interlocking Function 19.2 Ni-E has a useful function that provides various optical adjustments and settings by interlocking with switching of objectives or zoom operation. You can save time and efforts required for optical adjustments and settings that are needed when changing microscopy conditions.
  • Page 153 Chapter 3 Individual Operations <Field diaphragm: Example: Intelligent Setting> • Attach Address 3 of the motorized nosepiece: PlanApo20x NA 0.75, which is set to the field diaphragm [100%]. • Attach Address 4 of the motorized nosepiece: PlanApo60x NA 0.95, which is set to the field diaphragm [INTELLIGENT]. •...
  • Page 154 Chapter 3 Individual Operations Setting items Interlocking Conditions for interlocked operation  ON (with the button checked) Requires to connect a motorized nosepiece (Parfocal correction) Automatically corrects the difference (that requires defined objective information) between the focus position of and [FOCUS POS. SET] below should have objectives.

  • Page 155: Using The Ergo Controller

    Chapter 3 Individual Operations Using the Ergo Controller The ergo controller is connected to the Ni-E main body via a Operation panel connector box, and is used to control the motorized operation of the main body. Y knob Concurrent use with the joystick not available The ergo controller and the joystick cannot be used Z Speed XY Speed...
  • Page 156 Chapter 3 Individual Operations Using [FUNC SET] of the ergo controller on the DS-L3 [MICROSCOPE SETUP] screen allows you to change the default setting of the operation button function. Function Allocated to the CW/CCW Function Allocated to a Switch Other Than Button the CW/CCW Button Functions Allocatable to the CW/CCW Button and Other Buttons/Switches...
  • Page 157 Chapter 3 Individual Operations Using the Focus Knobs The focus knobs on the ergo controller are operated in the same manner as the focus knobs on the Ni-E main body. Pressing the Z speed switch on the operation panel of the ergo controller will switch the operation mode of the knobs between coarse, fine, and extra fine (with coarse being the factory setting).

  • Page 158: Using The Joystick Controller

    Chapter 3 Individual Operations Using the Joystick Controller The joystick is connected to the Ni-E main body via a connector box, and is used to operate the motorized XY stage. Connection cable XY Speed LED (to ERGO/JOY in the connector box) Constant Speed switch Z Speed LED Coarse...

  • Page 159: Capturing Images

    Chapter 3 Individual Operations Capturing Images A microscope image can be captured with a DS camera head DS camera attached to a tube or zooming port. head Use DS-L3 or DS-U3 DS Camera Control Unit. DS-L3 is connected with the USB connector or DSC1 connector at the back of the microscope or the DSC2 connector on the connector box to control the camera.
  • Page 160 Chapter 3 Individual Operations Photomicroscopy The photomicroscopy procedure is described below. See the instruction manual bundled with the DS-U3 or DS-L3, or the instruction manual for the microscope's setup/control software “Ni Setup Tool” for details including the camera settings. Using DS-L3 You must choose at least the following information: ...
  • Page 161 Adjust the camera’s white balance. To adjust the white balance, press the WB button while capturing an image of a clear section of the specimen slide. (For fluorescent photomicrography, Nikon recommends adjusting the white balance under normal bright-field microscopy conditions before capturing images.)
  • Page 162 Chapter 3 Individual Operations ■ Manual DSC zooming port Work with the zoom operation with the zoom dial on the right of the zooming port. The dial is indexed in 0.2x increments. Use these indexes for reference in making adjustments. The dial will click at 0.6x, 1x, 1.5x, and 2x magnification.
  • Page 163 Even if the overall magnification is the same on the monitor, the combination of objective and camera zoom can result in significant variations in exposure time. Nikon recommends increasing the magnification of the objective rather than of the zoom. (Generally, the numerical aperture of the objective increases with magnification. The larger the numerical aperture, the brighter the resulting image.)

  • Page 164: Operation On Ds-L3

    Chapter 3 Individual Operations Operation on DS-L3 23.1 Setting Up the Microscope If you use the microscope as it is set up as default, a specific setup is not required, however, if you want to change the setting to meet your needs, you want to use useful functions, or you want to set the information on optical elements, use DS-L3 for the operation below.
  • Page 165 Chapter 3 Individual Operations Shown below is the actual settings. See (1) through (7). Saving the setting After completing the setting, be sure to press the [SAVE] button on the [MICROSCOPE SETUP MENU] screen to save the settings. If DS-L3 is turned off without saving settings, previous values are restored.
  • Page 166 Chapter 3 Individual Operations (2) Configuring the Device Connection Information (See DS-L3 Chapter 6 “3 Setting the Connections of Motorized Units”.) (2-1) Connecting the digital camera (See DS-L3 Chapter 6 “3.1 Configuring the Connection of Digital Camera”.) Perform this configuration when a camera control unit is connected to the main body using a trigger cable to capture image by sending trigger signal from the DSC connector on the main body.
  • Page 167 Chapter 3 Individual Operations (3) Configuring the Button Functions (See DS-L3 Chapter 6 “4 Configuring the Functions of Buttons”.) [SETUP MENU] → [BUTTON FUNC] Change the function of buttons equipped with the Ni-E microscope and the ergo controller. The function of buttons located on the DS-L3 screen can be selected as desired here.
  • Page 168 Chapter 3 Individual Operations Units Supported for Buttons Operation Button Target Unit [INTSL] Shutter built in the HG precentered fiber illuminator [FL TURRET] Shutter built in the epi-fluorescence cube turret on the 1st layer (default setting) [FL 2nd] Shutter built in the epi-fluorescence cube turret on the 2nd layer FL SHUTTER Button [EPI/DIA/AUX] Motorized shutter connected to the EPI SHUTTER connector at the position set in “(2)
  • Page 169 Chapter 3 Individual Operations ■ Enabling operation buttons →(See DS-L3 Chapter 6 “4.2.5 Enabling/Disabling the Button Operation”.) Enable/disable the buttons located on the front and lateral right/left on the base of the Ni-E microscope. The buttons can only be configured as a group (front, lateral left, lateral right) not individually.
  • Page 170 Chapter 3 Individual Operations ■ SLEEP Button (See DS-L3 Chapter 6 “4.1.2 Showing/Hiding the [SLEEP] Button”.) Set to show or hide the [SLEEP] button on the [MICROSCOPE CONTROL] screen to turn ON/OFF the sleep mode on DS-L3. The sleep state is the state where the power supply to motorized devices is stopped to minimize the generation of noise.
  • Page 171 Chapter 3 Individual Operations (4) Configuring the Movement (See DS-L3 Chapter 6 “5 Configuring the Movement of Motorized Devices”.) Set the movement of the Ni-E microscope and motorized [SETUP MENU] → [MOVEMENT] devices. (4-1) (4-2) (4-3) (4-4) (4-5) Configuring the movement (4-1) Configuring Interlocking (See DS-L3 Chapter 6 “5.1 Configuring Interlocked Operation”.)
  • Page 172 Chapter 3 Individual Operations Set the focus position to interlock with ALF (Parfocal Correction). Press the [FOCUS POS. SET] button before specifying the objective on the optical path, and focus it and press the [SET] button. Notes on settings Set all addresses of the nosepiece. Set the same focus position as the one for the previous objective at a position where no objectives are attached.
  • Page 173 Chapter 3 Individual Operations ■ CAPTURE INTERLOCK (See DS-L3 Chapter 6 “5.1.7 Configuring the Interlocked Operation with Capture Command Sending or Trigger Signal Output”.) Specify to interlock the optical path of a motorized quadrocular tilting tube and a filter cube with an image capture using the [CAPTURE] button on the microscope main body.
  • Page 174 Chapter 3 Individual Operations (4-3) Z Position Nosepiece Rotation Stop (See DS-L3 Chapter 6 “5.3 Disabling the Rotation of the Motorized Nosepiece Depending on the Position of the Elevating Section”.) This function is to prevent the objective from contacting the specimen. When the elevating section is above than the designated position, the nosepiece rotation can be disabled.
  • Page 175 Chapter 3 Individual Operations (4-4) Nosepiece Reverse Rotation (See DS-L3 Chapter 6 “5.4 Disabling the Reverse Rotation of the Nosepiece”.) Since the focal depth of the objective with low magnification is deep, the objective may get too closer to a specimen. If the objective is switched to a high magnification objective in this case, the tip of the objective may contact the specimen.
  • Page 176 Chapter 3 Individual Operations (5) Set Mode Function (See DS-L3 Chapter 5 “2 Using MODE”.) In order to use the MODE function, “register” the motorized devices for the modes and “save” the status of the registered motorized device. “Load” the saved mode on the [MICROSCOPY CONTROL] screen. See Chapter 3 “19.1 MODE”...
  • Page 177 Chapter 3 Individual Operations (6) Configuring Other Functions (See DS-L3 Chapter 6 “6 Configuring Other Functions”.) (6-1) Ni-E Display Panel (See DS-L3 Chapter 6 “6.1 Setting the Display of the Ni-E Front Display Panel”.) Set a display pattern to be displayed on the display panel of Ni-E microscope at the system startup. The display pattern depends on the type of the motorized unit connected and is inclusive of 9 patterns.
  • Page 178 Chapter 3 Individual Operations (6-4) Software Limit (See DS-L3 Chapter 6 “6.4 Setting the Software Limits”.) Programmatically set the limit position of the microscope's elevating section and the motorized XY stage movement. In order to set the limit in XY direction, the motorized XY stage must be connected. Setting items Default setting Configurable range...
  • Page 179 Chapter 3 Individual Operations (7) Maintenance (7-1) FL Turret Drive Speed (See DS-L3 Chapter 6 “6.5 Setting the Driving Speed of the Epi-Fluorescence Cube Turret”.) Set the drive speed of the epi-fluorescence cube turret to High or Low. When a thick dichroic mirror is used for a filter cube, set the drive speed to “Low”. If you are using a two-layered epi-fluorescence cube turret, configure 1st and 2nd layers separately.

  • Page 180: Controlling The Microscope

    (1) What in Ni-E can be controlled by DS-L3 Required setting Device Operation available on DS-L3 (Reference in DS-L3 Instruction Manual) ECLIPSE Ni-E main body ON/OFF and lamp brightness control of None dia-illumination Adjustment of DIA field diaphragm Retraction and restoration...
  • Page 181 Chapter 3 Individual Operations (2) Buttons on the [MICROSCOPE CONTROL] Screen In order to control the microscope from DS-L3, press the [MIC] or [CAM-MIC] button on the [MICROSCOPE CONTROL] top screen to display the [MICROSCOPE CONTROL] or [CAM-MIC CONTROL] screen. The same operation is available from either screen, but the [MICROSCOPE CONTROL] screen is used as an example to summarize the operation in this document.
  • Page 182 Chapter 3 Individual Operations (3) Controlling Ni-E with buttons Pressing a button allows you to control the following parts of Ni-E or motorized devices: See the list for the function and descriptions on the buttons. (EPI: for epi-illumination) A. ST OP F.
  • Page 183 Chapter 3 Individual Operations Reference for Buttons with their Functions and Operation Procedures Operation Operation Button Function Reference in DS-L3 Instruction Manual Chapter 5, “1.1 Switching Switch the objective. the Objective (Motorized Nosepiece)” [NOSEPIECE] [Objective (Address)] Switch the optical path of the motorized quadrocular tilting tube to the binocular section.
  • Page 184 Chapter 3 Individual Operations Operation Operation Button Function Reference in DS-L3 Instruction Manual Chapter 5 “1.10 Adjusting Adjust the aperture diaphragm the DIA Aperture Diaphragm diameter of a motorized universal (Motorized Universal condenser. Condenser)” [A. STOP] Switch the ND of the HG precentered fiber illuminator.
  • Page 185 Chapter 3 Individual Operations Operation Operation Button Function Reference in DS-L3 Instruction Manual Open/close the built-in shutter in Chapter 5 “1.12.1 the HG precentered fiber Opening/closing the illuminator. (Equivalent to the Motorized HG Precentered [SHUTTER INTSL] button on the Fiber Illuminator's Built-in [SHUTTER INTSL] [INTSL] sub screen.) Shutter”...
  • Page 186 Chapter 3 Individual Operations Operation Operation Button Function Reference in DS-L3 Instruction Manual Chapter 5, “3 Entering the Enter the sleep state to reduce Sleep State (Noise noise. Reduction)” [SLEEP] Chapter 5 “4 Changing the Change the background color of Background Color of the the [MICROSCOPE CONTROL] [MICROSCOPE CONTROL]...

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