IGC

Leica DM LB2 | Reservations

  • Leica DM LB2
  • The Leica DM LB2 is an upright microscope for experiments with Brightfield and Differential interference contrast (DIC). DIC is especially suited for observation of highly transparent (unstained) specimens such as cells attached to Petri-dishes, thin tissue sections, or small embryos.

    Location : This micro is currently located at the Histology Unit

    Microscope  : Leica DM LB2

    Year Installed: 2003

    Camera: LEICA DFC250 (color, 2Megapixel)

    Optics : Brightfield and DIC

    NOTES :

    Keywords: widefield, color camera, upright microscope


Suggestion for description in "Materials and Methods":

Images were acquired on a Leica DM LB2 upright microscope, equiped with a Leica DFC250 color CCD camera, using the a 40x 0.75NA objetive and DIC optics.

Maintenance log:

  • 31-07-2014 - New computer installed to replaced old one. Compatible with the new IDS camera.
  • 16-04-2014 - The Wallaston prism for 40x objective.
  • 01-01-2014 - New temporary camera to replace the malfunctioning one.

Booking Calendar details

Configuration
Value
Booking type
Regular reservation
Responsible
Unidade Imagiologia
Minimum booking time (in min)
30
Schedule tolerance (in days)
7
Detele tolerance (in hours)
1
Booking restriction status
Disabled

Available objectives

Place mouse over Magnification to see the objective.
Magnification
Objective type
NA
Immersion
Working distance (mm)
Resolution @525nm in XY (µm)
old Pixel size IDS (µm)
old Pixel size TIS (µm)
Pixel size Leica DFC (µm)
DIC
Link
4x
HI PLAN
0.10
-
18
3.20
2.16
0.87
1.37
No
10x
HI PLAN CY
0.25
-
17.6
1.28
0.86
0.35
0.55
Yes
20x
HC PL FLUOTAR
0.50
-
0.18
0.64
0.43
0.24
0.27
Yes
40x
HCX PL FLUOTAR
0.75
-
0.40
0.43
0.215
0.09
0.14
Yes
100x
HCX PLAN APO
1.35
OIL
0.09
0.24
0.086
0.03
0.055
Yes

Setup

Turning system ON

  1. Turn on the microscope and adjust settings for brightfield and DIC (see instructions below)

    on/off switch

  2. Log on into the computer using your Agendo credentials.

  3. Place sample in microscope and set up as:

  4. Initialize/Open current camera by clicking in the green play icon.

    uEye open camera


Turning OFF

  • If you used any immersion lenses be sure to clean them.
  • TAKE OUT EVERYTHING YOU BROUGHT!, eg, pipettes, slides, etc...

Transmitted light - Brigthfield (FOR DIC SEE BELOW!)

  1. Set the upper filter wheel to the first positionm ("1").

    imt2_upper_filter_wheel

  2. Pull out both the upper Analiser (left) and Wallaston prism trays (Right).

    dmlb2_bf_slids

  3. Set the lower filter wheel (on condenser) to the “BF” and for magnifications above 4x confirm that the front lens of the condenser is in the light path (ie, slide this lens out of the light path only for magnifications of 4x or lower!).

    dmlb2_lower_filter_wheeldmbl2_light_ring_slide

    • Focus on your sample and confirm that you have proper koehler ilumination (see below)

      IF YOU DO NOT KNOW HOW TO DO THIS, ASK FOR ASSISTANCE!

  4. Focus the image with the coarse and fine drive. An additional focusing sensitivity can be switched on by moving the control from right to left.

    focus

  5. Adjust the lamp brightness according to your needs, but do this only after setting for Koehler illumination.

    brightness


Setting for Koehler illumination

  1. We advise you to start with the basic brightfield setup (see previous explanation).
  2. Insert into the light path the Analiser (upper) and Polariser (below condenser) trays; remove the Wallaston [Nomarski] prisms (both upper and lower). Look through eyepieces and rotate polariser for full extinction (darkness)

    dmlb2_dic_slids

    Analiser slid (top - below eyepieces)

    dmlb2_dic_polarizer

    Polariser tray (bottom - under the condenser)

  3. Re-insert Wallaston [Nomarski] prisms (both top and bottom) appropriate for the objective.

    NOTE: THE 40X AND 100X PRISMS ARE MISSING! UNTILL WE GET A REPLACEMENT DIC WITH THESE OBJECTIVES IS NOT POSSIBLE!

  4. Adjust contrast by rotating the knob in the upper Wallaston prism

Setting for Differential Interference Contrast (DIC)

  1. Make shure you have already a focused sample.
  2. Narrow completely the field diaphragm.

    dmlb2_field_diaphragm

  3. Form a sharp image of the edge of the diaphragm using the condenser height adjustment.

    imt2_condenser_heightkoehler

  4. Centre the image of the field diaphragm with the two centering screws.

    centering screws

  5. Open the field diaphragm, until it just disappears from the field of view. Adjust the aperture diaphragm until you obtain optimum contrast.

Note: Koehler illumination will not work with some objectives (ex: 2.5x and 100x).


Setting for Differential Interference Contrast (DIC)

  1. Make sure the microscope view is set to the CCD camera, pull outward the little rod on the left of the eyepieces.

    dmlb2_view_selector

  2. Start the Capture software available.
  3. Adjust the brightness on the microscope and the exposure time and color on the software until you are satisfied with your image (exposure might be in "auto").

    brightness

  4. Readjust the focus on the microscope until the image appear optimally sharp on the software.
  5. For more advanced acquisition options, like Color correction or image size ajustment, please speak with the UIC personnel.

Extra Info

Transmitted light - Brigthfield

Differential Interference Contrast (DIC)