Histology Spring 2009 Faculty Dr. John Bigbee (Course Director) Dr. Jack Haar Dr. Randall Merchant Dr. Peter Boling

Histology Spring 2009 Histology runs from January 5th to April 20th Histology is closely integrated with Physiology, lectures usually precede Physiology Lecture and practical components are weighted approximately equal: Lecture: Multiple choice exams, required material is contained in the the syllabus and lecture Laboratory: Self-study using the Digital Histology program; computer-based, multiple choice exams 5 exams each with a written and practical component; exams #3, #4 and #5 combined written and practical One microscope laboratory/clinical correlation (Aging Organs, skin)

TOTAL PRACTICAL POINTS 117 TOTAL POINTS 255 + 2 (lab) TOTAL LECTURE POINTS 138 TOTAL PRACTICAL POINTS 117   TOTAL POINTS 255 + 2 (lab) Each lecture hour will have 4 questions each and each lecture topic will have approximately 6 practical questions each. All questions are worth one point. Exam #1: 80 points 31.1% of total points Exam #2: 50 points 19.5% of total points Exam #3: 48 points 18.7% of total points Exam #4: 28 points 10.9% of total points Exam #5: 49 points 19.0% of total points

Recommended textbook HISTOLOGY: A TEXT AND ATLAS Ross, MH and Pawlina,W 5th edition, 2006

Histology “Laboratory” No traditional microscope laboratory. Laboratory is entirely self-study, using the Digital Histology program (PC and Mac formats). All practical exams are administered on computers in the CBIL and will use images similar to those used in Digital Histology. Required structures are listed in the syllabus following each lecture outline. Practical exams will include both identification and functional questions. If you need help, come see us !!!!!!

Practice practicals

Digital Histology Copyright to Digital Histology is retained by its authors and VCU. The program is marketed globally, along with a companion book, by textbook publisher Wiley-Liss. The publisher has granted us a special dispensation to distribute it freely to VCU students, but you may not share it with others. This program is also governed by the Terms of Use for the VCU School of Medicine eCurriculum.

Histology Objective is to teach the normal microscopic anatomy of the tissues and organs of the body. Appreciation of pathology begins with an understanding of normal structure. Histology helps develop your powers of observation.

Microscopy concepts

Specimen preparation Fixation Dehydration Infiltration and Embedding Sectioning Staining Microscopy

Fixation - Prevents autolysis (proteolysis) Preserves cellular structure Minimizes redistribution of proteins and organelles C H O CH2 Formaldehyde Glutaraldehyde Fixatives: Protein fixatives (cross linking) Formaldehyde Glutaraldehyde Lipid fixative Osmium tetroxide

Conventional staining Light microscopy Hematoxylin (blue) (basophilia) Eosin (pink-orange) H & E Electron microscopy Lead citrate Uranyl acetate Electron dense (dark) Electron lucent (light)

(Periodic acid-Schiff, PAS) Histochemistry Detection of specific chemical groups Carbohydrates (Periodic acid-Schiff, PAS) Lipids (Osmium)

Artifacts “Any feature of a tissue section which is present as a result of tissue processing” Shrinkage Spaces due to extraction of cellular contents Precipitates Redistributed organelles Mechanical consequences of sectioning, e.g, folds

Fixation artifacts Living cell Poor fixation

The compound light microscope

LM-EM Comparison

Electron microscopy Transmission electron microscopy (TEM) Scanning electron microscopy (SEM)

Microscopy While an important property of a microscope is the ability to magnify an image, the more important feature is the capacity to resolve the details in the image. Resolution: The smallest distance of separation at which two objects can still be distinguished as individual structures. Based on the wavelength of the illumination source. Limits of resolution: Light microscope 200 nm Electron microscope 1 nm (membranes = 7.5 – 10 nm)

Resolution and Magnification Magnified and resolved (EM) Magnified but not resolved (LM) Magnification: Enlargement of the image; adds no new information Light microscope 4x-1000x Electron microscope 1000x-500,000x

Size of microscopic structures Millimeter (mm) = 1/1000 Meter, 10-3M Regions or portions of organs Micrometer (m) = 1/1000 mm, 10-3mm, 10-6M Cells and large organelles, e.g. mitochondria Nanometer (nm) = 1/1000 mm, 10-3 m, 10-9M Cell membranes, filaments Angström unit (A) = 1/10 nm, 10-10M Macromolecules

The virtual slide A digital image of microscopic section which is fully navigable and zoomifiable (magnifiable). Closely approximates the use of a microscope. Digitally capture contiguous small portions of the section at high magnification and then combine all these images into a single seamless image of the entire tissue section.

Making a virtual slide

Making a virtual slide Sun, et al., 2006. J. Microscopy, 224:158

Using the virtual slide image acquisition Computer assembly of image and file compression Storage on server 500 MB per image

Using the virtual slide Mikula, S, et al., 2007. Neuroimage, 35:9-15

https://java.vcu.edu/som-histology/

Virtual slide collection Cells and Tissues Cell/Mitosis Whitefish blastula Muscle tissue Cardiac muscle Skeletal muscle Connective tissue Ground bone Nervous tissue Peripheral nerve (ls) Peripheral nerve (xs) Spinal cord Eye Retina Organ systems Cardiovascular Integumentary Heart Thick skin Medium A&V Small A&V Lymphoid Lymph node Digestive Spleen Esophagus Thymus Fundic stomach Duodenum Reproductive Ileum Ovary Colon Uterus, secretory Submandibular gland Testis Liver Placenta Pancreas Respiratory Endocrine Trachea Pituitary Lung Thyroid Adrenal Urinary Kidney

Planes of section Cross section (cs) or transverse section (ts) Longitudinal section (ls) Oblique section

To increase screen image size On the desktop, right click then “Properties” Choose “settings” tab Adjust the slider to 800 x 600 screen resolution Click OK and Apply Use the same steps to change resolution back to normal

Practical strategies Go through the section with the list of required structures at the end of each syllabus outline. Go through the section with the “labels off” feature selected. Go through each section in random mode with “labels off” feature. Take individual section quiz for each topic. Compile sections and randomize with “labels off”. Take practice practical listed under self-assessments on eCurriculum.