Olympus BX51

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PRESENTATION

BX51

Olympus BX51 manual fluorescence microscope with mercury fluorescence illuminator, and Nomarski/DIC Prism for Transmited Light


Room: 2.1.15

IP: 10.101.134.44 (why?)


TBA


PosMagNAIMCorr.WD (mm)Glass (mm)Color CodeObs
14x0.13AirUPlanFL17--
210x0.40Air / WaterUPlanApo3.10--
320x0.20AirLCPlanFL7.55 - 6.75--
440x0.85AirUPlanApo0.20--
5100x1.25OilPlanC N---

Pos Position

Mag Magnification

NA Numerical Aperture

IM Immersion Medium

Corr. Correction Type

WD Working Distance

Glass Coverglass

Obs Observations


PositionExcitation (nm)Emission (nm)FluorophoresModel
U-MWU2BP 330-385LP >420DAPI, hoechst-
U-MNIBBP 470-490LP >515GFP, Alexa488, cy2, FITC-
U-MNGBP 530-550LP >590RFP, YFP, Alexa546, Alexa568, cy3, TRITC-
U-MSWBBP 420-480LP >515only w/ cromossina-


Transmitted light

Halogen lamp

Fluorescence

Olympus U-RFL-T Mercury lamp


Axiocam  mRm, 1.3MPixels, monochromatic cooled CCD – acquisition with uManager

TIS DFK 1.9MP Sony CCD color camera – acquisition with IC Capture software or uManager


ACCESSORIES


 

BASIC PROCEDURE FOR COLOR IMAGES

1.Turn on the microscope transmitted light illumination at the power switch, and adjust light intensity on the right hand side round knob. Push the eyepiece/camera selector pole all the way in so that you can see through the eyepieces. Choose desired objective and bring your sample into focus.

2.Turn on PC, and start IC Capture 2.4 program.

3.Pull the eyepiece/camera selector all the way out. You should see your sample on the viewing window. If not, press button 1. Make sure FPS are at 15.00 (button 3).

4.Press button 2 to visualize the histogram (easier: grayscale, visualize zeros, log scale options), and adjust EXPOSURE (button 4) until you see histogram peaks close to, but not at, 255 (pure white / saturation). UNLESS YOU KNOW WHAT YOU’RE DOING and wish to do pre-processing, leave Brightness, Contrast, Gain, and Sharpness at ZERO (uncheck all “Auto” cheboxes), and Gamma, Auto Reference, and Color Balance at default values.

5. Using the onscreen image, set up Koehler Illumination, and readjust your exposure, if necessary. Koehler illumination usually has to be repeated for each objective.

6.When you’re happy with your image, right-click on it and pick “Save Image” from the menu. Save in your folder.

7.When you’re done with work, turn to the lowest magnification objective, adjust the light intensity knob to zero, and switch off the microscope. Copy your images from Voxel 6, and switch off the acquisition computer.

Olympus BX51 is also equipped with DIC and can acquire fluorescence images with a black&white Zeiss AxioCam mRm camera. If you need these, please ask Telmo or Luís for assistance.


IC Capture 2.4

uManager


LINKS / USER MANUALS


EQUIPMENT RESPONSIBLE

Telmo gabriela_rodrigues

Telmo Nunes

FCUL

Microscopy technician

mevarrimento@fc.ul.pt

Room 2.1.15

Ext: 22181

Gabriela Rodrigues

FCUL / CE3C

Assistant professor

mgrodrigues@fc.ul.pt

Room 2.3.10

Ext: 22310