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Leica DMIRB Inverted Microscope with Color & B/W Cooled Digital Cameras

Location: 6129 Thurston Bowles

 

Notices

Contents


Introduction

The Leica microscope is a standard inverted microscope set up for transmitted light, differential interference contrast (DIC or Nomarski) and fluorescence microscopy.  Images may be viewed by eye or viewed and saved to computer with either of 2 cameras.  For transmitted light and very bright fluorescence imaging a chilled color camera is mounted on top of the trinocular and can be used to capture high resolution digital images.  For dimly fluorescent samples or photometric quantification, a cooled B/W CCD camera (an HQ2) made by Photometrics is attached to the side port on the left of the microscope.  DIC images may also be captured using either camera, although the color camera can capture with higher resolution.  Darkfield is also available forthe 40x & 63x oil immersion objectives.  The specific hardware attached to this microscopy is listed below.

The Equipment

The Microscope:
  • Leica inverted DMIRB microscope
     
  • Objectives:
Objectives
Mag. NA type WD Adjustable Collar cover slip Immersion part no. Objective Wollaston *** Condenser Wollaston 
5X * 0.11 Plan ~9 mm - #1.5 air   20 B1
10x * 0.3 Fluor 6 mm - #1.5 air   20 B1
20x 0.4   long corr 0 - 2.0 mm air   20 C or D
40x 0.55   long corr 0 - 2.0 mm air   40 C or D
40x ** 1.0 to 0.5 Plan Fluor 240 um aperture #1.5 oil   40 C or D
100x ** 1.4 to 0.7 PlanApo 90 um aperture #1.5 oil   100 C or D

Use only Leica immersion oil with oil immersion objectives
* These objectives prefer a number #1.5 (~170 um thick) glass cover slip.
** Suitable only for imaging though cover slips due to small working distance
*** When not using DIC, the objective Wollaston should be in the blank (H) position
corr - cover slip adjustment should be set for maximum clarity with fine high contrast objects or set the dot on the dial on objective to the corresponding objective thickness. e.g. for #1.5 (170um) cover slips dot should be at 0.2 (you'll have to interpolate), or e.g. standard culture dishes (1.0 mm thickness) set dot to 1.0

 

 

Cameras:

Comparison of the Cameras attached to the Leica Inverted Microscope

Camera
Comparison

Monochrome

Color

Camera HQ2 OrcaER MicroPublisher 3.3 (sometimes 5.0)
Manufacturer Photometrics Hamamatsu Q-Imaging
Optimized for: Dim fluorescence &/or quantification of fluorescence intensity.  B/W transmitted light imaging Dim fluorescence &/or quantification of fluorescence intensity.  B/W transmitted light imaging High resolution transmitted & bright fluorescence
Location on microscope On left side port On left side port On top if trinocular
Light transmission from microscope 100% 100% 100% or 50%
Computer to use \\Lister (computer on the right) \\Lister (computer on the right) \\Snell (computer on the left)
Resolution Better than presented by 0.63x coupler Better than presented by 0.63x coupler Better matched to image presented by microscope
Preview Computer monitor (C-Imaging) Computer monitor (C-Imaging) Computer monitor (C-Imaging or Q-Capture)
Max..field of view size 1300 by 1024 pixels 1300 by 1024 pixels 2048 by 1542 pixels
Min. field of view size   Sub region of interest supported 512 by 385 pixels
Intensity depth 8 or 12 bit B/W 8 or 12 bit B/W 8 or 10 bit each RGB channels (24 or 30 bit total)
Color balance not applicable not applicable Good after performing auto-balance
Sensitivity good good mediocre
Far capability good good none
Quantification of intensity excellent excellent mediocre
Acquisition software C-Imaging C-Imaging C-Imaging or Q-Capture or ImageJ
Cooled yes yes no
Interface Firewire 1394 Firewire 1394 Firewire 1394
Power up Any time, before C-Imaging is run Any time, before C-Imaging is run Can connect/disconnect with power on
Power source External Power brick (switched)/ External Power supply Firewire cable

Computer:


Parts of the Leica DMIRB microscope     

For printing 

                      

Setting Up the Microscope


Image Acquisition

Acquiring Images Using SimplePCI   <--- please click on this link  


Acquiring Images Using the Q-Imaging Software with the MicroPublisher Color Camera


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Copyright 2001-2018  Dr. M. Chua, School of Medicine, University of North Carolina, Chapel Hill, NC 27599
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Last Updated: 2015-12-28