In the past decades imaging technologies are increasingly used to model the dynamics and structure of biological systems. Biomedical imaging is now an integral part of biological and medical sciences and is used in both clinical practice and research. In this session some of the latest imaging technologies were reviewed.

1. Sima Salahshor, BSc, MSc, PMP, PhD
Department of Laboratory of Medicine & Pathobiology,
Faculty of Medicine, University of Toronto
& ScienceHA, Inc.
Lecture part of the “Cellular Imaging in Pathobiology” course
Course ID #LMP1006H (http://www.lmp.utoronto.ca/course/lmp1006h)

2. Full body
scan,
Whole animal
scan
Organ,
Tissue
imaging
Cellular
imaging
Chromosome
Genes
DNA
Molecular
imaging
Single
molecule
Radiology, Sonography,
Radioiodine whole body scan
Ultrasound, Spectroscopy
MRI, CT scan, PET scan
(General diagnostic imaging)
FISH, SKY
C-banding, G-banding
Light-, Confocal-, Two-photon-, Fluorescence microscopy
Electron microscopy (SEM, TEM)
FLIM, FRAP, TIR-FM , AFM
FRET, X-ray crystallography,
Super-resolution microscopy,
Laser tweezers,
Raman spectroscopy
Imaging Technologies

3.  Whole body MRI scan
 Imaging of Multiple Myeloma and Related
Plasma Cell Dyscrasias
 Full body scanner (detect
object under clothing)
 Backscatter X-ray systems
 Millimeter wave technology
Employs non-ionizing submillimeter microwave
radiation (reflects extremely high frequency radio
waves off the body)
Reference: Ronald C. Walker et al
J Nucl Med July 1, 2012 vol. 53 no. 7 1091-1101
http://jnm.snmjournals.org/content/53/7/1091.full.pd
f+html
Health effect and safety of backscatter x-ray or
millimeter wave scanners: Not fully investigated!
Reference: http://www.iacrs-rp.org/

4. Small-animal imaging
Reference:
Hoffman J M , Gambhir S S Radiology 2007;244:39-47
A. whole-body micro-PET image
B. micro-CT image
C. micro-SPECT image
D. Optical reflectance fluorescence image
E. T2-weighted micro–MR image
F. Optical bioluminescence image

5.  MRI Magnetic Resonance Imaging
 uses a powerful magnetic field, radio frequency pulses and a computer to produce detailed
pictures of organs, soft tissues, bone and virtually all other internal body structures.
 CT X-ray Computed Tomography
 combines special x-ray equipment with sophisticated computers to produce multiple images or
pictures of the inside of the body.
 PET Positron Emission Tomography
 uses small amounts of radioactive material to diagnose or treat a variety of diseases, including
many types of cancers, heart disease and certain other abnormalities within the body.
 SPECT Single-Photon Emission Computed Tomography
 a special type of computed tomography (CT) scan in which a small amount of a radioactive drug is
injected into a vein and a scanner is used to make detailed images of areas inside the body where
the radioactive material is taken up by the cells. SPECT can give information about blood flow to
tissues and chemical reactions (metabolism) in the body.
 Ultrasound
 exposing part of the body to high-frequency sound waves to produce pictures of the inside of the
body.
 Optical Imaging
 the image formed by the light rays from a self-luminous or an illuminated object that traverse an
optical system.

6. The Compound Light Microscope
The Stereo Microscope (dissecting microscope)
The Electron Microscope
The Scanning Probe Microscope (SPM)

7. An electron microscope is a type of microscope that produces an electronically
magnified image of a specimen for detailed observation.
The electron microscope (EM) uses a particle beam of electrons to illuminate the
specimen and create a magnified image of it.
 TEMTransmission Electron Microscope
 can achieve magnifications of up to 50,000,000x
 SEMScanning Electron Microscope
 magnification up to 5,000,000x
Both SEM and TEM are useful in biology and geology, as well as in materials science.

8. Reference:
Girard F, Batisson I, Frankel GM, Harel J and Fairbrother JM (2005). Interaction of enteropathogenic and
Shiga-Toxin producing Escherichia coli with porcine intestinal mucosa: role of Intimin and Tir in adherence.
Infection and Immunity 73: 6011

9. Floral primordia of Allium sativum (garlic) captured with the Epi-Illumination technique.
Credit: Dr. Somayeh Naghiloo
University of Tabriz, Department of Plant Biology, Faculty of Natural Sciences
http://www.nikonsmallworld.com/galleries/photo || Small World, 2012 Nikon competition winner

10. Mouse C2C12 cells in prophase imaged with 3D
structured illumination microscopy (SIM).
Condensed chromosomes are stained with DAPI
(red), the nuclear lamina and microtubuli are
immunolabeled with an anti-lamin B (blue) and an
anti-tubulin antibody (green), respectively.
Reference:
Lothar Schermelleh, Ludwig-Maximilians-Universität München http://www.cell.com/cell_picture_show-superres

11. Imaging by WFM, LSCM and 3DSIM
10 µm section of mouse small intestine, fixed in
formaldehyde, cryosectioned, stained with DAPI (blue),
anti-tubulin (red) and phalloidin (green) and mounted in
glycerol stained with DAPI.
A: WFM recorded on a Leica fluorescence microscope
with a Hamamatsu Orca CCD camera.
B: LSCM on a Zeiss 710 microscope.
C: 3DSIM recorded on an OMX microscope.
Images courtesy of Paul Appleton and Emma King.
Reference:
Innovation in biological microscopy: current status and future
directions. Bioessays. 2012 May;34(5):333-40 by Swedlow JR.
WMF =A widefield Microscope (one type of Fluorescence Microscope)
LSCM = Laser Scanning Confocal Microscopy
3D-SIM = Structured Illumination Microscopy (super-resolution technique).
SIM can image up to 10 microns past the coverslip into the sample.

12. A powerful instrument for
microbiological Investigation.
AFM has the advantage of
imaging almost any type of
surface, including polymers,
ceramics, composites, glass,
and biological samples. AFM
provides a 3D profile of the
surface on a nanoscale.
Reference:
Atomic force microscopy: a nanoscopic view of microbial cell surfaces. Micron. 2012
Dec;43(12):1312-22 by Dorobantu LS, Goss GG, Burrell RE.

13.  Karyotype
 Chromosome analysis gives you a full picture of the structure of each chromosome as
well as the number of chromosomes present in each cell.
 FISH Fluorescent In Situ Hybridization
 FISH analysis doesn't give you a picture of each chromosome but it does tell you how
many copies of a certain chromosome are present in each cell.
 SKY Spectral Karyotyping
 Permits the simultaneous visualization of each human or mouse chromosome in a
different color, facilitating the identification of chromosomal aberrations.
 CGH Comparative Genomic Hybridization
 Utilizes the hybridization of differentially labeled tumor and reference
DNA to generate a map of DNA copy number changes in tumor genomes.

14. Reference: H.-U.G. WEIER, J. KWAN1, C.-M. LU, Y. ITO, M.
WANG, A. BAUMGARTNER ,S.W. HAYWARD6 J.F. WEIER,
H.F. ZITZELSBERGER
Credit: Cancer Genomics Program,
Departments of Pathology and Oncology, University of Cambridge

15. Superresolution Microscopy
Photoactivated localization microscopy (PALM)
Stochastic optical reconstruction microscopy (STORM)
Near-field scanning optical microscopy (NSOM)
Stimulated emission depletion microscopy (STED)
Saturated structured-illumination microscopy (SSIM)
Ground state depletion (GSD) microscopy
two-photon laser-scanning microscopy (TPLSM).
The study of lymphocyte activation
requires observation of samples that
vary in size over six orders of
magnitude. This figure shows the T cell
receptor (TCR)-mediated signalling
pathway and microscopy techniques
used at three levels of sample size.
Reference:
Imaging techniques for assaying lymphocyte
activation in action. Lakshmi Balagopalan, Eilon
Sherman, Valarie A. Barr & Lawrence E.
Samelson

16. Applications of imaging techniques
Reference:
Lakshmi Balagopalan, Eilon Sherman, Valarie A. Barr & Lawrence E. SamelsonImaging techniques for assaying lymphocyte activation in action

17. Reference:
Cancer imaging by optical coherence tomography: preclinical progress and clinical potential.
Vakoc BJ, Fukumura D, Jain RK, Bouma BE.
Overview of intravital imaging approaches in
preclinical cancer research

18. Imaging Technique Spatial Resolution Key Use
Multi-photon Microscopy 15 – 1000 nm Visualization of cell structures
Atomic Force Microscopy 10 – 20 nm Mapping cell surface
Electron Microscopy ~5 nm Discerning protein structure
Ultrasound 50 μm Vascular imaging
CT/MicroCT 12 – 50 μm Lung and bone tumor imaging
MRI/MicroMRI 4 – 100 μm Anatomical imaging
fMRI ~1 mm Functional imaging of brain activity
MRS ~2 mm Detection of metabolites
PET/MicroPET 1 – 2 mm Metabolic imaging
The various micro versions of the imaging modalities (MicroCT, MicroMRI, MicroPET) as well as the
microscopy techniques (Fluorescence, Multi-photon, Atomic, Electron) are primarily used in either cellular or
animal studies. The remaining modalities (Ultrasound, CT, MRI, MRS, PET) are more widely used clinically.
Reference:
Kherlopian et al. BMC Systems Biology 2008, 2:74
Comparison of imaging technology for
systems biology

19. No. 1: Magnetic resonance imaging (MRI) & Computed Tomography (CT) scan
No. 5: Functional Magnetic Resonance Imaging (fMRI)
No. 8: Molecular Breast Imaging (MBI)
Medical imaging technology has revolutionized health care.
MRI CT scan fMRI MBI
Reference:
Ann Tracy Mueller, 2013, http://bit.ly/1QOkYgr

20. Email: s.salahshor@utoronto.ca || URL: http://bit.ly/1hTp48a
Twitter: @SSalahshor
For more information about LMP1006H course visit:
http://www.lmp.utoronto.ca/course/lmp1006h