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CMN Imaging Core 

January 16, 2013

 

1 

LEICA DM IRE2 MICROSCOPE MANUAL 

 
Neuroscience Imaging Core 
Rightmire Hall 
Ohio State University 
 
Director: 

Tony Brown

 

Rightmire 060 
292-1205 

[email protected]

 

 
Facility Manager: 

Paula Monsma 

Rightmire 062 
292-3025 

[email protected]

 

 
This manual prepared by Tony Brown. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Summary of Contents for DMIRE2

Page 1: ...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 mon...

Page 2: ...brightness 13 To focus using the focusing knobs 14 Using the focusing buttons 15 Changing the coarseness of the focusing 16 Eyepieces 17 Placing a slide on the Z galvo stage 17 Transmitted light detec...

Page 3: ...control panel Laser scan head with 3 fluorescence detectors Condenser lens Side port Focus knob Eyepieces Transmitted light detector Z galvo stage detachable Halogen lamp intensity adjustment wheel Fi...

Page 4: ...RIGHT Halogen lamp housing transmitted light Mercury lamp housing epi fluorescence Focus knob X Y stage movement Filter holders empty Transmitted light detector selection knob Condenser turret Tube le...

Page 5: ...VE TURRET DETAILED VIEW objective turret objective prism turret filter cube turret cuvesfou DIC analyzer Epi fluorescence field diaphragm controls area of illumination Epifluorescence Illumination dia...

Page 6: ...rism 63x water immersion 100x oil immersion E DIC objective prism 40x oil immersion 63x oil immersion CONDENSER TURRET POSITIONS Condenser turret position Description Matching objectives BF Empty for...

Page 7: ...luorochromes RED red fluorochromes CFP far red fluorochromes SCAN empty slot select port VIS light to eyepieces SIDE light to scanner BOTTOM disabled switch tube lens disabled LCD display panel button...

Page 8: ...t Indicates current step size setting for focusing arrow indicates upper limit has been set arrow indicates lower limit has been set Indicates current filter cube Indicates current objective Indicates...

Page 9: ...with If you accidentally press this button you will see the word EXIT flashing Press the LEARN button again to exit the learn mode Learn mode DO NOT USE Set lower limit of travel for objective DO NOT...

Page 10: ...n fluorochromes RED cube for red fluorochromes CFP cube for far red fluorochromes or cube for CFP see Paula for details SCAN empty slot Notes The cube in the CFP position is normally the far red cube...

Page 11: ...11 Changing objectives Use the objective turret control buttons on the left side of the microscope Current objective indicated on LCD display panel Press upper key to increase magnification Press low...

Page 12: ...y objective or the chance of mixing of different immersion media the microscope will not allow you to move freely between these modes using the objective turret control buttons To switch from one mode...

Page 13: ...ically 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 po...

Page 14: ...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...

Page 15: ...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 buttons The only way to focus above the upper limit is to use t...

Page 16: ...settings Setting Step size S0 Fine S1 Medium fine S2 Medium coarse S3 Coarse You can use any step size with any objective but when you first select an objective the default step size will be as follow...

Page 17: ...ece so that the image appears in focus to your left eye Close the left eye and adjust the right eyepiece so that the image appears in focus to your right eye The eyepieces are now parfocal Placing a s...

Page 18: ...ition 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 rotate c...

Page 19: ...r focusing knob until the image of the aperture is sharp If necessary center the field diaphragm in the field of view using the two centering screws located on the front of the condenser Open field ap...

Page 20: ...turret to select phase ring PH1 for 10x PH2 for 40x Focus on specimen Insert the Bertrand lens into the optical path using by rotating the tube lens module from SCAN to B Focus on the phase ring using...

Page 21: ...ret An objective prism in the objective prism turret To set up DIC Select a DIC objective any of the immersion objectives The objective DIC prism required for that objective is displayed on the micros...

Page 22: ...movement in Z axis for acquisition of Z stacks Universal stage holder holds a wider range of dishes To remove Z galvo stage Unscrew the two thumbail screws circled in red Place stage still attached t...

Page 23: ...y If you use too much oil it may run down the side of the objective and damage the optics To apply oil to an objective dip oil applicator in immersion oil bottle allow excess to drain off apply oil to...

Page 24: ...sh area of the lens tissue for each blot Repeat blotting until no more oil comes off onto the lens tissue Wipe the metal housing around the lens using lens tissue It typically takes 2 or 3 sheets of l...

Page 25: ...ly opposite in the objective turret from the objective you want to apply oil to i e 3 positions away from the current objective Objective you want to use Objective in opposite position 100x oil 20x mu...

Page 26: ...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...

Page 27: ...ectives or any other optics Never use any liquid cleaners or solvents to clean the microscope optics or any part of the microscope When to ask for Paula s assistance Changing mercury or halogen bulbs...

Page 28: ...CMN Imaging Core January 16 2013 28...

Page 29: ...Finest 63 100x S1 40x S2 20x S3 10x SC 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 bot...

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