BSCI
427 FALL,
2004
PRINCIPLES OF MICROSCOPY S.
WOLNIAK
LECTURE
SCHEDULE
Dates Topic Readings*
Aug. 30 Introduction
to the light microscope M
(ch. 1, 2, 4), R (1-6, 23-24)
Kohler
illumination, bright field S
(1-31), D (ch. 1,3), Ob
microscopy
Sept. 1,8 Image
formation; diffraction M
(ch. 5), R (7-11), S (1-31)
D
(ch. 1)
Sept. 13 Resolution,
numerical aperture M
(ch. 4, 6), R (7-22)
objective
lens design S
(1-31,63-69)
D
(ch. 1)
Sept. 15-22 (Digital)
Photography and photomicrography, M
(ch. 3,13) R (113-120)
Filters,
light temp., chromatic S
(70-72),
aberration D
(ch.4-10)
Sept. 27,29 Phase
contrast; darkfield microscopy M
(ch. 7),R (25-27,30-31,
80-87,91-92), S
(32-38)
Oct. 4,6 Fluorescence
microscopy, microfluoro- M
(ch. 11), R (27-30,88-90)
metry,
microspectrophotometry; S
(40-45)
light
detectors Z
or L or Of
Oct. 11 Polarized
Light Microscopy M
(ch. 8,9), R (32-36,94)
Birefringence,
Compensation, BR S
(46-53), Bennett
Oct. 13 Differential
Interference Contrast M
(ch. 10), R(36-44,92-94)
Hoffman
Modulation Contrast S
(46-53),
Interference
Reflection Microscopy Hoffman
Interference
Microscopy S
(54-61)
Oct.
18 NO
CLASS
Oct. 20 Digital microscopy and computer
image M
(ch. 13-15), R (121-125), P
analysis
Oct.
25-Nov. 3 (Confocal)
Scanning optical microscopy M
(ch. 12), R (44-49,96-98)
Deconvolution
Microscopy Handbook,
Wright, Stelzer,
Petroll
Nov.
8 Lasers
for microbeam surgery,
fluorescence,
optical traps Block
Acoustic
microscopy M
(137-158)
Nov.
10 Exam
Nov. 1- Dec. 11 Training
with the 510 and DV microscopes
0107
Microbiology (sign up for microscope use)
Dec.
13 10:30-12:30
Oral Presentations/Picture Analysis
___________
* Readings: M
= Murphy, D.B.
2001. Fundamentals of Light Microscopy and
Electronic
Imaging, Wiley
R
= Rawlins, D.J.
1992. Light Microscopy. Bios Scientific Publishers
S = Spencer, M. 1982. Fundamentals
of Light Microscopy
D
= Delly, J.G. 1988. Photography through the Microscope.
Eastman
Kodak Co.
L = Leitz booklet on fluorescence
microscopy
Z = Zeiss booklet on fluorescence
microscopy
Of = Olympus booklet on
fluorescence microscopy
Ob = Olympus booklet on
brightfield microscopy
P = Photometrics booklet on CCD cameras
Block
= article on
optical tweezers
Bennett = article on polarized light
microscopy
Stelzer = article on confocal scanning light microscopy
Wright
= article on
confocal scanning light microscopy
Petroll = article on confocal scanning
light microscopy
BSCI
427 (2 credits) FALL,
2003
PRINCIPLES OF MICROSCOPY S.
WOLNIAK
2204
HJP
Office
hours: By
appointment - call 5-1605 or email at sw36@umail.umd.edu
Course
Requirements
November
10th This hourly exam
will cover the theoretical aspects of imaging through the light
microscope.
In room 0107 of the
Microbiology Building, we have two new microscopes that represent the best in
confocal instrumentation and digital image processing/deconvolution. This
course provides you with an unusual opportunity to learn how to use these
microscopes, and I plan the lecture cycle of this course so that you can spend
some time using these instruments.
Step 1: training with
a simple specimen. Dr. Bob Brown
will train you in the basics of confocal imaging with the Zeiss 510 and with
the DeltaVision Image Reconstruction Microscopes. You will start with a
standard, stable specimen to generate a set of color images that will enable
you to see what is possible. Save and transfer these files to the workstations,
because you can use them in the presentation (step 3, below), especially if
your cells prove to be troublesome.
Step 2: imaging some
of your own material. After you
have been checked out on the microscopes, you need to speak with me about
strategies to get good confocal or deconvolution images with your specimens.
Then, you need to able to sign up for some time with one or both of the
microscopes in room 0107 Microbiology. Take several micrographs of representative
cells and a few sets of image stacks. Try to obtain multi-color images, and/or
superimposed fluorescence and transmitted light images (DIC or phase contrast).
Transfer the files to the workstations, so that you can manipulate the images
without having to sign up for more microscope time. Translate the files to
TIFF, so that they can be viewed with Adobe Photoshop. We can burn CDs from the workstations so that
you have permanent records of your work, and so that you can show the others in
the class what you have been able to generate in a few short hours.
Step 3: presentation
time. We will meet at class time
on Monday, December 13th, in room 0107 Microbiology to look at
everyoneÕs images. At this stage, it is more important to produce images of
cells than to generate masterpieces that will be preserved in museums. If
images have flaws, it is actually better, because we can use this presentation
class meeting to discuss ways to reduce problems and improve the likelihood
that you can get cover pictures and color plates that will convey structural
information accurately and beautifully.
From November 1st
onward, Bob Brown and I will be available to help you in getting trained to
generate images of your cells.