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LEICA DM IRE2 MICROSCOPE MANUAL
Neuroscience Imaging Core
Rightmire Hall
Ohio State University
Director: Tony Brown
Rightmire 060
292-1205
brown.2302@osu.edu
Facility Manager: Paula Monsma
Rightmire 062
292-3025
monsma.1@osu.edu
This manual prepared by Tony Brown.
1
CMN Imaging Core
January 16, 2013
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Summary of Contents for Leica DM IRE2
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Page 1: Leica Dm Ire2 Microscope Manual
LEICA DM IRE2 MICROSCOPE MANUAL Neuroscience Imaging Core Rightmire Hall Ohio State University Director: Tony Brown Rightmire 060 292-1205 brown.2302@osu.edu Facility Manager: Paula Monsma Rightmire 062 292-3025 monsma.1@osu.edu This manual prepared by Tony Brown. CMN Imaging Core January 16, 2013... -
Page 2: Table Of Contents
TABLE OF CONTENTS LEICA DM IRE2 MICROSCOPE MANUAL ........ 1 Getting to know the microscope ..........3 Microscope control panel ............7 LCD display panel ..............8 LCD display panel buttons ............9 Changing filters ............... 10 Changing objectives..............11 To switch between dry and immersion objectives .... -
Page 3: Getting To Know The Microscope
Getting to know the microscope VIEW FROM LEFT Transmitted light Transmitted illumination column Eyepieces light detector (tilts back for better access to stage) Condenser lens Z- galvo stage (detachable) Filter cube turret Laser scan head with 3 fluorescence detectors Side port Focus knob Halogen lamp intensity... - Page 4 VIEW FROM RIGHT Halogen lamp housing (transmitted light) Filter holders (empty) Transmitted light detector selection knob Condenser turret Mercury lamp housing (epi-fluorescence) Objective turret Tube lens module with Bertrand lens X-Y stage Focus knob movement CMN Imaging Core January 16, 2013...
- Page 5 OBJECTIVE TURRET (DETAILED VIEW) objective objective turret prism turret Epifluorescence Illumination diaphragm (controls brightness of illumination) Epi-fluorescence field diaphragm (controls area of illumination) DIC analyzer filter cube turret cuvesfou CMN Imaging Core January 16, 2013...
- Page 6 OBJECTIVE PRISM TURRET POSITIONS Objective Description Matching objectives turret position Empty (for bright field or phase contrast) DIC objective prism 20x multi-immersion 63x water DIC objective prism immersion 100x oil immersion 40x oil immersion DIC objective prism 63x oil immersion CONDENSER TURRET POSITIONS Condenser Description...
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Page 7: Microscope Control Panel
Microscope control panel LCD display LCD display panel buttons Red light shows current filter selection switch filter cube GFP: green fluorochromes RED: red fluorochromes CFP/: far red fluorochromes SCAN=empty slot select port VIS=light to eyepieces SIDE=light to scanner BOTTOM: disabled Mercury lamp switch tube lens shutter... -
Page 8: Lcd Display Panel
LCD display panel arrow arrow Indicates Indicates current indicates indicates position of step size lower limit upper limit objective setting for has been has been relative to focusing upper limit Indicates Indicates current current filter objective cube D= dry mode I= immersion mode Indicates Indicates... -
Page 9: Lcd Display Panel Buttons
LCD display panel buttons Set lower Set upper Adjust step Learn mode Toggle (DO NOT between limit of limit of size for voltage USE!) travel for travel for coarseness readout objective objective of focusing (DO NOT (DO NOT objective USE!) USE!) readout Do not press the LEARN button (this will enter you... -
Page 10: Changing Filters
Changing filters Use the motorized fluorescent filter cube changer on the microscope control panel: Press left button Press right button to rotate filter to rotate filter cube turret left cube turret right Red light shows current filter cube selection The filter cube turret contains three filters plus an empty slot ... -
Page 11: Changing Objectives
Changing objectives Use the objective turret control buttons on the left side of the microscope Press upper key to increase magnification Press lower key to decrease magnification Current objective indicated on LCD display panel CMN Imaging Core January 16, 2013... -
Page 12: To Switch Between Dry And Immersion Objectives
To switch between dry and immersion objectives The standard objectives on our microscope are grouped into three blocks or modes: 100x oil immersion oil immersion mode 63x oil immersion 40x oil immersion 20x oil/water/glycerol immersion multi immersion mode 40x dry dry mode 10x dry To reduce the chance of immersion medium getting... -
Page 13: Adjust Halogen Lamp Brightness
Adjust halogen lamp brightness Use the dial on the front left side of the microscope stand near the base Halogen lamp brightness control The lamp voltage will display automatically on the microscope control panel display when the intensity dial is adjusted To switch off transmitted light illumination adjust lamp intensity to 2.5V then continue rotating dial beyond this point... -
Page 14: To Focus Using The Focusing Knobs
To focus using the focusing knobs One way to focus is using the focusing knobs located on the left and right side of the microscope. Turning the knob so that your thumb moves away from you focuses down Turning the knob so that your thumb moves toward from you focuses up Focusing knobs (turn in direction of arrows to... -
Page 15: Using The Focusing Buttons
Using the focusing buttons Another way to focus is to use the focusing buttons located on the right side of the microscope Press upper focus key to focus up Press lower focus key to focus down Note about upper limits: If an upper limit is set for the objective (see LCD display panel) then the objective will not move above that limit if you are focusing using the focusing... -
Page 16: Changing The Coarseness Of The Focusing
Changing the coarseness of the focusing The focusing is electronic and has five settings: Setting Step size Fine Medium fine Medium coarse Coarse You can use any step size with any objective, but when you first select an objective the default step size will be as follows: Objective Setting... -
Page 17: Eyepieces
Eyepieces Adjusting interpupillary distance: Adjust eyepieces to match interpupillary distance by moving the eyepieces closer together or further apart Adjusting parfocality Focus on specimen using electronic focusing controls Close right eye and adjust the left eyepiece so that the image appears in focus to your left eye ... -
Page 18: Transmitted Light Detector Selection Knob
Transmitted light detector selection knob For any transmitted light observation (bright field, DIC, phase contrast), this knob should be in the vis position. Bright field observation Bright field observation means observation with transmitted light using no contrast enhancement method (e.g. no phase contrast or DIC) For bright field observation: ... -
Page 19: Koehler Illumination
Koehler illumination Koehler illumination is essential to obtain good transmitted light images Select objective Open the condenser aperture diaphragm (move lever to the right) Focus on specimen Close the field aperture diaphragm (move lever to left) ... -
Page 20: Phase Contrast
Phase contrast For phase contrast, you need: a phase contrast objective a matching phase ring in the condenser turret To set up phase contrast: Select a phase contrast objective (i.e. either of the dry objectives) rotate objective prism turret to BF (empty) position ... -
Page 21: Differential Interference Contrast (Dic)
Differential interference contrast (DIC) For DIC, you need: A DIC objective (i.e. any of the immersion objectives) A polarizer above the condenser (no need to insert this – it is kept in the light path at all times) ... -
Page 22: Switching Stage Holders
Switching stage holders The microscope has two stage holders: Z- galvo stage (permits rapid and precise movement in Z axis for acquisition of Z stacks) Universal stage holder (holds a wider range of dishes) To remove Z-galvo stage: ... -
Page 23: Using Immersion Objectives
Apply oil to the objective or to the coverslip before placing your slide on the stage Use only LEICA IMMERSION OIL Use the minimum amount of oil necessary If you use too much oil it may run down the side of... -
Page 24: Cleaning Objectives
Cleaning objectives This microscope is equipped with the highest quality objectives, with a total value exceeding $30,000! You must exercise great care to preserve these objectives To clean oil off immersion objectives: Remove specimen from stage Blot (not wipe!) off excess oil using lens tissue ... -
Page 25: Applying Oil To Objective Without Removing Slide
Applying oil to objective without removing slide FOR ADVANCED USERS ONLY! You can use the following procedure if you are observing your slide and you wish to switch to an oil immersion objective without removing the slide from the stage holder. ... -
Page 26: Specimen Preparation
Specimen preparation Some general tips: Select fluorochromes that are optimally excited by the confocal laser lines For multiple labeling, the less overlap between the excitation and emission spectra the better Always use #1.5 coverslips Mount coverslip to slide securely and seal with nail enamel or mounting medium that solidifies ... -
Page 27: Dos And Don'ts
Adding or removing filter cubes Using the 63x water immersion objective Cleaning oil off 10x and 40x dry objectives Using the Leica objective warmer Using the Bioptechs heated chamber Any other time you are unsure what you are... - Page 28 CMN Imaging Core January 16, 2013...
- Page 29 Extras Setting Step size Objective Finest 63,100x Coarsest Note about the coarsest focusing setting (SC): Press both upper and lower focus keys simultaneously to switch to the coarsest focusing setting (SC) Press both focus keys simultaneously to switch back to fine focusing setting (S0-S3) CMN Imaging Core January 16, 2013...