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Olympus BX63 upright microscope

P-G Desmarais Building, Room 5308-2
Advanced Microscope Tier 1 usage price
Instrument awarded to Dr Marina Martinez by the Canadian Foundation for Innovation (CFI)

 

Applications

  • Widefield
    • Brightfield
    • Polarized light
    • DIC

    • Fluorescence

  • Color camera


Light sources

  • LED lamp for transmitted light

  • X-Cite NOVEM-S XT910 for fluorescence

SourceExcitation (nm)FluorophoresNominal power (mW)Measured power (mW)
365

DAPI, Hoechst

50081
435
CFP480
475
FITC, GFP41067
500-600-
[500-600]
3150
509/22
[498-520]		YFP

554/23
[542-565]		TRITC
143
578/21
[567-588]		mCherry

635


Cy5

45543

735


Cy7

40038

Objectives

  1. 4x/0.16 Air

  2. 10x/0.4 Air

  3. 20x/0.75 Air

  4. 40x/0.95 Air

  5. 60x/1.35 Oil

  6. 100x/1.4 Oil

  7. Empty

PositionNameBrandFull nameIDMagnificationNumerical apertureImmersionWorking distance (mm)Transmittance
(% [nm])		TechniqueCoverslip thickness (mm)
14x/0.16
Air		Olympus4x/0.16 Air
UPlanSApo

UPLXAPO4X.pdf

4x0.16Air16

>90% at 550nm

BF, Fluo-
210x/0.4
Air		Olympus10x/0.4 Air
UPlanSApo		UPLXAPO10X.pdf10x0.4Air3.1

~90% at 550nm

BF, Pol, DIC, Fluo0.17
3

20x/0.75
Air

Olympus20x/0.75 Air
UPlanSApo

UPLXAPO20X.pdf

20x0.75

Air

0.6

>90% at 550nm

BF, Pol, DIC, Fluo0.17
4

40x/0.95
Air

Olympus40x/0.95 Air
UPlanSApo

UPLXAPO40X.pdf

40x

0.95

Air

0.18

~90% at 550nm

BF, Pol, DIC, FluoAdjustable 0.11-0.23
560x/1.35
Oil		Olympus60x/1.35 Oil
UPlanSApo

UPLXAPO60XO.pdf

60x1.35
Oil
0.15

~90% at 550nm

BF, Pol, DIC, Fluo0.17
6

100x/1.4 Oil

Olympus100x/1.4 Oil 
UPlanSApo

UPLXAPO100XO.pdf

100x1.4

Oil

0.13

~90% at 550nm

BF, Pol, DIC, Fluo0.17
7

Empty










BF: Brightfield
Pol: Polarized light
DIC: Differential interference contrast
Fluo: Fluorescence

Filters

  1. Empty
  2. DAPI
  3. FITC
  4. TRITC
  5. mCherry
  6. DAPI/FITC/TRITC/Cy5/Cy7
  7. Empty
  8. U-FDICT DIC Analyzer (Nomarski prism)
PositionNameBrandIDExcitation FilterDichroic mirrorEmission FilterComments
1Empty





2DAPIOlympusU-FUNA360-370 410LP420-460 

U-FUNA

3FITCOlympusU-FBNA470-495 505LP510-550 

U-FBNA

4TRITCOlympusU-FGWA530-550 570LP575-625 

U-FGWA

5mCherryOlympusU-FYW540-585 595LP600LP

U-FYW

6DAPI/FITC/TRITC/Cy5/Cy7

LED DA/

FI/TR/CY5/C

Y7-5X-A-000

5 filters within the Novem

  • FF01-387/11

  • FF02-485/20

  • FF01-560/25 

  • FF01-650/13

  •  FF01-740/13

FF408/504/581/667/762-Di01 FF01-440/521/607/694/809
7Empty





8

DIC Analyzer

OlympusU-FDICT DIC---DIC Analyzer

Detector

  • Olympus DP80

    Camera

    		Olympus DP80

    Sensor Type

    		CCD

    Sensor Category

    		Color/Monochrome

    Nb Pixels

    		 1.45 M
    12.5 M (pixel shift)

    Pixel Layout

    		1360 x 1024
    4080 x 3072 (pixel shift)

    Pixel size

    		6.45 um

    Sensor size

    		8.8 mm x 6.6 mm 

    Sensor diameter

    		11 mm

    Bit depth

    		14-bit
    Speed at full resolution7.7 images/s

    Max QE

    		55 %
    Readout noise7 e⁻

    Cooling

    		-10 C Pelletier

    Dark Current

    		0.4 e⁻/pixel/sec

    Full well capacity

    		17 000 e-

    Dynamic Range

    		1:2300

    Interface

    		USB 3.0

    Mount

    		B4 mount (2/3 inch Bayonet)

    Olympus_DP80 Camera_User Guide.pdf
    Olympus_DP80_Brochure.pdf

  1. If not already done, turn on the computer (#1) and log in to Windows using your UdeM credentials
  2. Remove the dust cover from the microscope
  3. Turn on the microscope power bar (#2)

  4. Turn on the microscope touchscreen (switch is located at the back on the right of the touch screen) (#3)

  5. When using the instrument for the first time, it is necessary to import the microscope configuration before starting the software. See the First Use section below.

  6. Start-up CellSens Dimension
  7. Wait for the software to open (yes it's long...)

When using the instrument for the first time, it is necessary to import the microscope configuration into the software. This procedure is usually carried out during the training session. However, it can also be used to reset the software if it does not display correctly, for example. 

Running this procedure will erase all your experiment protocols and reset the software to its original settings (ask for support if you are not sure).

  1. If open, close CellSens Dimension and wait until it is completely closed (up to 30 seconds)
  2. On your Desktop open the Softwares folder
  3. Double-click BX63 CellSens Settings

  4. A script will run and a black window will appear briefly

  5. When the message Settings for CellSens have been imported successfully appears, click OK to close it

  6. A script will run and a black window will briefly appear
  7. You can now reopen CellSens Dimension
  8. Wait for the software to open (yes it's long...)

During this procedure, you will:

  • Set the microscope in a safe configuration

  • Load your sample

  • Find and adjust the focus

Once completed, your sample will be ready for acquisition.

  • If this has not been done yet, select the lowest-magnification objective, 4×.

  • Press 4x to select the 4x objective

    The 4× objective is the safest to use due to its long working distance (16 mm). The sample will appear in sharp focus well before the objective gets close. It is recommended to always focus first using the safest objective. Since the objectives are parafocal, focusing with these objectives will make it easier to locate the sample when switching to higher-magnification objectives.

  • Press Escape to raise the lenses to their highest position

  • Place the test slide on the microscope stage with the coverslip toward the objective

Using a test slide will significantly reduce the time needed to set up the instrument.
  • If necessary, move the stage so that the sample is centered under the objective
  • Select the desired imaging mode (Brightfield, or fluorescence, DAPI, FITC, TRITC...)
  • Adjust the focus using the main focus knob while looking through the eyepieces until the image is perfectly sharp

The focus is around Z = 19000 um. The Z position value is visible on the touch screen.

  • Select Off to turn off the illumination

First perform the initial focusing with the safest objective before selecting a higher-magnification objective.

After completing the initial focus:

  • Select the desired air objective (10×, 20×, or 40×)

The 40× objective is the best air objective because it has the highest numerical aperture (0.95).

  • Select the desired imaging mode (Brightfield, or fluorescence, DAPI, FITC, TRITC...)
  • Adjust the focus using the fine focus knob while looking through the eyepieces until the image is perfectly sharp
  • Select Off to turn off the illumination

Your sample is ready for acquisition!

After completing the initial focus:

  • Select the desired oil objective (60× or 100×)

The 100× objective is the best air objective because it has the highest numerical aperture (1.4).

  • The microscope will automatically raise the objectives so that the sample is accessible

  • Place a drop of oil on your sample

  • Click OK

  • The microscope will automatically lower the objective to the previous location
  • Select the desired imaging mode (Brightfield, or fluorescence, DAPI, FITC, TRITC...)
  • Adjust the focus using the fine focus knob while looking through the eyepieces until the image is perfectly sharp
  • Select Off to turn off the illumination

Your sample is ready for acquisition!

  • Files can be saved temporarily (during acquisition) on the local C: drive (desktop)
  • At the end of each session, copy your data to your external drive and delete it from the local C: drive
  • You can store your files on the D: drive (Data Storage). If you do, please create a folder per laboratory using the principal investigator last name. Within, create one folder per user (Firstname_Lastname).

In any case, your files should be removed from the C: drive.

  • Save your data
  • Close CellSens Dimension
  • Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
  • If used, clean oil objectives with lens cleaner and paper
  • Select the 4x objective and place the objectives in a safe position (Escape)
  • Turn off the microscope power bar (#2)
  • Turn off the computer
  • Cover the instrument with the protective dust cover
  • Take back your samples including ones in the microscope
  • Leave the microscope and the working area clean
  • QC
  • Complete installation setup
  • Scrip for CellSens personalization
  • System relocated to Room 5308-2
  • Added to Faces
  • XCite NOVEM-S Installed

Stand

  • Olympus BX63 Serial 0G00080

Condenser

  • Motorized condenser

Camera

  • Olympus DP80 Serial 8K48090

Stage

  • Motorized stage SHe Serial 127808

Workstation

  • HP Z4 G4 Workstation
  • I155446-CIB
  • Intel Xeon W-2102 @ 2.9 GHz
  • Motherboard HP Z4 G4 Chipset Intel C422
  • RAM 32 GB DDR4 1200 MHz (4 x 8 GB)
  • OS 1 TB SSD 550 MB/s
  • 2 TB HD Data Storage (2 x 1 TB spanned volume) 162 MB/s
  • Video Card nVidia Quadro P620 2 GB DDR5 1.39 TFlops
  • Monitor Dell P2415Q display 24' 3840 x 2160 
  • Software CellSens Dimension v3.2 ID SKPX-KN7D-YER5-CPSA

Isolation table

  • None

Consumables

Troubleshooting

Please ensure that the microscope touchscreen is turned on. The switch is not easily accessible and is located on the back at the right of the touchscreen pad.

FAQ

No. This is an upright microscope designed to look at specimen mounted between a slide and a coverslip

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Content published under CC BY-SA 4.0. Sharing allowed under the same license with attribution: "Original content by l'Institut Courtois d'innovation biomédicale, used under CC BY-SA 4.0"