North Seattle Community College
Spring 2012
NANO230
Confocal and Optical Microscopy
Laser Scanning Confocal Microscope
Compound Light Microscope
Introduction:
Laser Scanning Confocal Microscopy (LSCM) is a powerful raster imaging
method, which can resolve features down to ~200 μm. The optical path of an
LSCM is shown below:
LSCM’s illuminate the specimen with a laser light source. The laser is scanned
over the specimen, and the resulting fluoresce is focused thru a pinhole aperture
on to a photomultiplier tube (PMT) detector.
Experimental:
In this exercise you will image the same specimen in both the confocal
microscope and the compound light microscope. You will need to hand in a 1-2
page color electronic report which contains the following:
1. LSCM image with scale bar
2. Compound light microscope with scale bar
3. Specimen description (very short paragraph), size of field, magnification, NA,
transmission medium for each image
4. Paragraph which explains how the images are different and why. Specifically
address color, and depth of field.