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Motic AE30-31 Instruction Manual
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Motic AE30-31 Instruction Manual

Episcopic-fluorescence attachment ef-inv-ll inverted microscope
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Episcopic-Fluorescence Attachment EF-INV-ll
Inverted Microscope AE30-31
Instruction Manual
MOTIC INCORPORATION LTD.
Prepared by Motic Instruments Inc. Canada

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Summary of Contents for Motic AE30-31

  • Page 1 Episcopic-Fluorescence Attachment EF-INV-ll Inverted Microscope AE30-31 Instruction Manual MOTIC INCORPORATION LTD. Prepared by Motic Instruments Inc. Canada...
  • Page 2 This instruction Manual has been prepared for users of the AE-INV-ll Epi-Fluorescence Attachment used in conjunction with the Motic AE30/31 Inverted Microscope. We are constantly endeavouring to improve our instruments and to adapt them to the requirements of modern research techniques and testing methods.
  • Page 3 A filter designed to allow the passage of radiation of wavelengths shorter than a given limit. Motic Instruments Inc. Canada October 06, 2006...

  • Page 4: Table Of Contents

    Table of Contents Names of Component Parts Setting up the Instrument Assembly Microscopy Troubleshooting Table Care and Maintenance Motic Instruments Inc. Canada October 06, 2006...

  • Page 5: Names Of Component Parts

    Names of Component Parts Light source Epi-fluorescence attachment Fig. 1 Epi-fluorescence attachment EF-INV-ll Power supply unit Light protective shield Motic Instruments Inc. Canada October 06, 2006...

  • Page 6: Setting Up The Instrument

    Installing the Epi-fluorescence attachment Please see illustrations while assembling the attachment Required tools Allen hex keys: 1.5mm, 2.5mm and 5mm (supplied with attachment). Before Starting Turn off the microscope power switch and unplug the power cord Motic Instruments Inc. Canada October 06, 2006...
  • Page 7 Rear dust cover Microscope AC inlet See Figures 3a and 3b Remove side dust covers located on each side of the microscope base. Fig. 3a Fig. 3b Right side cover Left side cover Motic Instruments Inc. Canada October 06, 2006...
  • Page 8 Fix the main body sleeve in this position by using the three 5mm Allen hexagonal screws. Fig. 4 Epi-fluorescence attachment main body sleeve Notch facing left Motic Instruments Inc. Canada October 06, 2006...
  • Page 9 Epi-fluorescence horizontally into the sleeve until it cannot be pushed in any further. To secure, tighten the fixing screws using a hexagonal Allen key. Fig. 5 Shutter slider with Filter holder Motic Instruments Inc. Canada October 06, 2006...
  • Page 10 Light cut-off slider • To prevent the specimen from fading during temporary pauses in observation, slide the shutter to the left. • When using transmitted light illumination, slide the shutter to the left. Motic Instruments Inc. Canada October 06, 2006...
  • Page 11 • Using an Allen hex key, secure the cassette firmly with the two 2.5mm clamp screws. Fig. 7 2.5mm clamp screws Filter cassette Motic Instruments Inc. Canada October 06, 2006...
  • Page 12 Slot the two pins into the two matching holes on the microscope opening surround. • Using an Allen hex key, secure the filter cube cover retaining plate firmly with the two 2.5mm clamp screws. Fig. 8 Filter cube cover 2.5mm Clamp screws retaining plate Motic Instruments Inc. Canada October 06, 2006...
  • Page 13 Never perform lamp centering with the UV filter cube in the optical path as harmful UV radiation from the lamp may enter the eyes, possibly resulting in loss of vision. Fig. 9 Allen hex screws Filter cube clamp plate Motic Instruments Inc. Canada October 06, 2006...
  • Page 14 (Fig.10). Fig. 10 1.5mm Allen hex key Three filter cubes can be positioned on the filter slider. Fig. 11 Motic Instruments Inc. Canada October 06, 2006...
  • Page 15 Secure filter cube clamp plate with Allen hex screws (Fig. 9) Install and secure the filter cube cover on both the right and left sides with the knurled retaining screws. Fig. 12a Left cover Motic Instruments Inc. Canada October 06, 2006...
  • Page 16 (Fig. 12b) on the right cover. Fig.13 Insert the excitation ID tag (Supplied with the filter cube) Fasten the excitation method changeover lever to the filter cube Insert the filter cube ID tags Motic Instruments Inc. Canada October 06, 2006...
  • Page 17 Attach the light-shielding device to the eyepiece tube housing using the clamp screw on the right. • Adjust the height and angle of the light-shielding device with the knurled knob on the left. See Figure 14 Motic Instruments Inc. Canada October 06, 2006...

  • Page 18: Assembly

    Loosen the set screw at the base of the focusing knob with a 2.5mm Allen hex key. • While pulling out the collector lens focus knob, insert the collector lens into the lamp house in the direction indicated by insertion arrow. Motic Instruments Inc. Canada October 06, 2006...
  • Page 19 Tighten the set screw at the base of the focusing knob with a 2.5mm Allen hex key. • Tighten the cheese-head slotted screw, following lamp alignment and focusing. • Affix the rubber cap covering collector lens focus knob. Motic Instruments Inc. Canada October 06, 2006...
  • Page 20 Fig. 16b Turn the bayonet ring in the direction indicated by the arrow Secure the bayonet ring by turning it as far as possible in the direction of arrow. Motic Instruments Inc. Canada October 06, 2006...
  • Page 21 Insert the lamp into the lower flexible lamp clamp base and tighten the clamp screw. See Fig.18a • Insert the lamp into the upper lamp socket hole supported by the metal plate with the cooling fins and fasten the clamp screw. See Fig.18b Motic Instruments Inc. Canada October 06, 2006...
  • Page 22 Insert the bottom tab of lamp house cover over the inner metal surround of the lamp house and pivot the lamp socket cover to close. Secure with the clamp screw. • The safety cut-off micro switch will “click” as you secure the lamp house cover clamp screw. Motic Instruments Inc. Canada October 06, 2006...
  • Page 23 Plug the lamp input connector into the output connector on the rear panel of the power supply and secure by tightening the locking ring. • Plug in the power cord supplied with the power supply into the AC receptacle on the rear panel of the power supply. Motic Instruments Inc. Canada October 06, 2006...
  • Page 24 The “Run Time” counter displays the elapsed time POWER LAMP / READY LAMP indicator LAMP TIME RUN TIME counter 0. 0. 0. 0. Counter RESET PAD Reset IGNITION IGNITOR button POWER POWER switch Motic MHG-100B Mercury Lamp Power Supply Motic Instruments Inc. Canada October 06, 2006...
  • Page 25 When the lamp has been allowed to stabilize it gives off high intensity light concentrated at certain discrete wavelengths (e.g. 365, 400, 440, 546 and 580nm) making it an ideal source of illumination for fluorescence microscopy. Motic Instruments Inc. Canada October 06, 2006...
  • Page 26 The final scattering of the beam is checked on an actual fluorescent specimen. • The size of the image of the arc can be made bigger or smaller by manipulating the collector lens focus knob. Motic Instruments Inc. Canada October 06, 2006...
  • Page 27 Cutting out the excessive light is useful in preventing contrast from being diminished. Decreasing excessive illumination is useful in protecting the specimen from fading. Fig. 21 Field aperture diaphragm lever Epi-fluorescence attachment main body sleeve Shutter slider with Filter holder Epi-fluorescence attachment main body Motic Instruments Inc. Canada October 06, 2006...
  • Page 28 The shutter is also used to cut off the reflected light illumination when observing the specimen by transmitted light illumination (e.g. as in Phase contrast microscopy). Motic Instruments Inc. Canada October 06, 2006...
  • Page 29 Dichronic mirror is placed at 45º angle relative to the incoming excitation light, the dichroic mirror performs two functions: it reflects the shorter wavelength exciting radiation light to the specimen and transmits the longer wavelength fluorescence. Motic Instruments Inc. Canada October 06, 2006...

  • Page 30: Microscopy

    This is generally the case for fluorophores that generate a single emitting species in specimens with reasonably low levels of background auto fluorescence. Longpass Filter Motic Instruments Inc. Canada October 06, 2006...
  • Page 31 Block Block Block Block Pass Pass Narrow Bandpass Filter Wide Bandpass Filter FITC FITC TRITC Both FITC and TRITC are visible Only FITC is visible Motic Instruments Inc. Canada October 06, 2006...
  • Page 32 Oil immersion objectives are labelled with the additional engraving “Oil” and are to be immersed in oil between the specimen and the front of the objective. The immersion oil supplied by Motic is synthetic, non-fluorescing and non-resining oil, with a refractive index of 1.515 Normally, cover glass must be used with oil immersion objectives with a few exceptions.
  • Page 33 Electronic photomicrography utilizes electronic detection devices to acquire images, inserting an IR (infrared) barrier filter in the light path before the detection device may produce better results by avoiding erroneous readings caused by infrared light. Motic Instruments Inc. Canada October 06, 2006...

  • Page 34: Troubleshooting Table

    Field of view is partially obscured Revolving nosepiece not clicked into position. Shutter slider in intermediate position. Filter cube not installed correctly. Filter cube in intermediate position. Field diaphragm out of centre. Field diaphragm is stopped down Motic Instruments Inc. Canada October 06, 2006...
  • Page 35 Replace when necessary. Handle Multi- Band filters with great care as the very complex coatings wear easily. Special attention should be given to moisture. When filter is not in use, store in a desiccators. Motic Instruments Inc. Canada October 06, 2006...

  • Page 36: Care And Maintenance

    Proper handling of the microscope will ensure years of trouble free service. If repair becomes necessary, please contact your Motic agency or our Technical Service directly. The fluorescence vertical illuminator can accommodate three filter blocks and a dia-filter dummy block (devoid of filters) that enables normal brightfield observation. Filter blocks are mounted on a sliding rack and can be inserted into the optical path by means of a lever that is utilized to control the rack position.
  • Page 37 Endow GFP Bandpass Emission Set: Exciter HQ470/40x Dichroic Q495LP Emitter HQ525/50m This is the recommended filter set for the newer S65T GFP mutants. It is also recommended for wtGFP for which it was originally designed. Motic Instruments Inc. Canada October 06, 2006...
  • Page 38 Emitter HQ535/30m This set is designed to image YFP. It will not, in most cases, detect CFP when co-expressed in cells. By eye, fluorescence in the 535 nm pass-band will be seen as green. Motic Instruments Inc. Canada October 06, 2006...
  • Page 39 Arrangement of Filters in a Fluorescence Cube Inverted Microscope To Objective and Specimen Exciter Filter Dichroic Lamp Mirror HBO 100W Barrier Filter To Observation Method Motic Instruments Inc. Canada October 06, 2006...
  • Page 40 Most interference are mounted at a slight angle to allow for better imaging by suppressing ghost images. Emission filter D460/50m* The center wavelength is at 460nm; full band is 50nm [= ±25nm]. *m –emission Motic Instruments Inc. Canada October 06, 2006...
  • Page 41 (FRET) experiments. Green Glass – Longpass absorption glass from Schott Glassworks with cut-on wavelengths in the violet and blue-green regions A designation for high-performance filters with narrow cuton and cutoff wavelengths Motic Instruments Inc. Canada October 06, 2006...
  • Page 42 (See Figure Below) Signal to Noise Ratio Ratio of intensity of signal to that of the background. Stokes shift The distance between the peak absorption and emission of a dye, usually in nm. Motic Instruments Inc. Canada October 06, 2006...
  • Page 43 Shortpass and longpass filters, block or transmit wavelengths at specific cut-off wavelengths. Bandpass filters exhibit broadband or shortband transmission centered on a particular band of wavelengths. Filters performance is defined by the central wavelength (CWL) and by the full width at half maximum (FWHM). Motic Instruments Inc. Canada October 06, 2006...