This document provides an introduction to a histology course, including information about the instructor, course details, textbook, and an overview of histology as the study of tissues at the microscopic level. It describes the basic tissue types and techniques used to prepare tissue samples for microscopic examination, from biopsy to staining of slides. The goal of histology is to understand normal and diseased tissues at the cellular level in order to diagnose conditions like cancer and determine appropriate treatment.
This document provides an introduction to a histology course, including information about the instructor, course details, textbook, and an overview of histology as the study of tissues at the microscopic level. It describes the basic tissue types and techniques used to prepare tissue samples for microscopic examination, from biopsy to staining of slides. The goal of histology is to understand normal and diseased tissues at the cellular level in order to diagnose conditions like cancer and determine appropriate treatment.
This document provides an introduction to a histology course, including information about the instructor, course details, textbook, and an overview of histology as the study of tissues at the microscopic level. It describes the basic tissue types and techniques used to prepare tissue samples for microscopic examination, from biopsy to staining of slides. The goal of histology is to understand normal and diseased tissues at the cellular level in order to diagnose conditions like cancer and determine appropriate treatment.
This document provides an introduction to a histology course, including information about the instructor, course details, textbook, and an overview of histology as the study of tissues at the microscopic level. It describes the basic tissue types and techniques used to prepare tissue samples for microscopic examination, from biopsy to staining of slides. The goal of histology is to understand normal and diseased tissues at the cellular level in order to diagnose conditions like cancer and determine appropriate treatment.
Office Hours: Wed. 10 - 12 pm or by appt. (3161 HHB) Phone: 248-364-8689 email: [email protected]
Educational Background: Ph.D., Cell Biology, Michigan State University, M.B.A. Tarleton State University B.A., Biology, Western Michigan University C(ASCP), HTL(ASCP), Tarleton State University
Research: Dr. Telgenhoff’s research focuses on factors that affect wound healing. In particular, he is interested in the migration of skin cells into the wound area, and the effects of pathologic states such as diabetes and hyperlipidemia on this migration. His current studies utilizes cells grown in culture to mimic the wound environment in both normal and underlying pathologies.
Personal Interests: Woodworking, fishing, homebrewing, and travel
Course Description • Histology is the study of body organization and anatomy at the microscopic level. Simultaneous registration in the laboratory class (BIO 3141) is not required, but is encouraged. • In this class students will integrate cell biology, physiology, and pathology through a visual/structural approach to tissues and organ systems. • The textbook, combined with supplementary reading material, will be used to communicate and reinforce fundamental concepts in the areas of basic tissues and integrated cell biology and organ systems. • The format for the course includes conventional lectures as well as class discussion designed to promote active learning.
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Course Details • Class meets: T – TH 8 – 9:47am, 202 Dodge • Textbook (required): Ross MH & Pawlina W. 2011. Histology: A Text and Atlas • Prerequisite(s): BIO 1200 • Point Accumulation: Unit Exams (6 at 100 pts, lowest dropped): 500 Final Exam: 125 TOTAL 625
Our Text
Pawlina W. 2018
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What is Histology?
Cell
Organ System
Tissue Organ Use of images for educational purposes only
Four Tissue Types are the Building Blocks
of all Organs
1. Epithelial Tissue
2. Connective Tissue
3. Muscle Tissue
4. Neural Tissue
Use of images for educational purposes only
4 1/6/2020
How do we study cells, tissues, organs,
& organ systems? • Gross Level (Anatomy)
• Microscopic Level (Histology)
• Molecular Level (Cell Biology)
Use of images for educational purposes only
Histology Techniques • Many techniques are used to study cells and tissues.
• In this class, we will be using light microscopy
to observe sections of tissue mounted on glass slides.
• How are these sections prepared?
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Preparation of Tissues for Light Microscopy
1. Harvest tissue
2. Fixation to preserve structure of the tissue
3. Dehydration to remove water from the tissue
4. Clear the alcohol from the tissue
5. Infiltrate and Embed with Paraffin wax
6. Section tissue into thin slices (like deli meat)
7. Mount sections on a slide
8. Rehydration to prepare sections for staining
9. Stain to enhance contrast or highlight structures of interest
10. Observe with the light microscope
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• Is it cancer ?? • If it is… how serious is it? • Will I need treatment? What kind?
Next step in the journey….
• “We need to look at the tissues. We need a biopsy.” – Is it cancer or is it benign ? – Is it invasive? – What treatments?
• Histology: study of normal tissues • Pathology: study of diseased tissues • Tissue: – Building blocks of organs – group of cells working together to carry out a specific function • Biopsy: small bit of tissue removed from patient for examination with microscope
Limitations of histotechnique • Tissue is dead • Only a slice: 2D • Color artificial: – Purple nuclei – Pink cytoplasm • Need to know “what is normal”
What kinds of treatment?
Are hormone receptors present?
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From microscopy slide to Pathology Report:
POWER of Histotechnique Specimen: Clinical History 49 year old female who was found to have a mass on physical exam and mammography. Stereotactic core biopsy reveals malignancy. Clinical Diagnosis: Right breast cancer
Microscopic Description Slides A3- A4 consist of a portion of breast tissue in which is located an infiltrating ductal carcinoma of the breast. The malignancy is characterized by infiltrating nests of malignant cells in which there is glandular formation. The cells demonstrate a moderate degree of nuclear pleomorphism, with some of the cells having central nucleoli. However, the mitotic rate is less than 1 mitosis per 10 high power fields. Overall the carcinoma is grade II. Adjacent to the carcinoma are areas of ductal carcinona in situ. Histologically the tumor is 2mm from the inked margin. No lymphatic invasion is appreciated.
Diagnosis Infiltrating ductal carcinoma of the breast, Grade II
Foci of ductal carcinoma in situ Tumor is 2 mm from the inked margins Metastatic cancer in 2/12 lymph nodes. Markers The tumor is estrogen receptor positive The tumor is Her 2 neu positive
Histotechnology
Theory and Practice
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Histotechnology
When tissues are removed from the body, there is
rapid onset of action of degradative enzymes, which start the process of autolysis
Thus, the tissues need to be immediately
processed, perhaps to isolate cells or frozen in order to be studied, or fixed, in order to preserve them for study as archival material
Histotechnology Frozen tissues need storage space in either liquid nitrogen or in minus 80 degree freezers which take up space
Fixed tissues are then subjected to a process of
dehydration before being infiltrated with paraffin wax (at high temperatures) in order to be able to store them at room temperature for use as archival material
The fixatives used preserve morphology of the tissue
but can alter cell surface molecules
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WELL FIXED SMALL BOWEL AND POORLY FIXED SAMPLE
Processing of tissue :
Freeze for Process for EM
histology/histochemistry/ & use for Process into paraffin blocks immunohistochemistry
-Fix -Dehydrate OCT in Dry ice plastic mold -Infiltrate with xylene -Infiltrate with hot paraffin wax -Make blocks for sections -Store at room temperature
Frozen or paraffin tissue can then be sectioned for histology 3--30 micron sections
15 1/6/2020
MATERIALS NEEDED TO FREEZE TISSUE SAMPLES
MATERIALS NEEDED TO PROCESS FIXED TISSUE
Place fresh or fixed, trimmed, tissue into
cassettes to be processed into paraffin blocks for sectioning at room temperature
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Paraffin embedded tissues ready for sectioning onto glass slides
HISTO: HISTOLOGY SECTIONS FOR VIEWING UNDER THE MICROSCOPE, using
BRIGHTFIELD illumination
Always review sections using the basic hematoxylin and eosin (H&E) stain before proceeding to perform a special stain
H&E= hematoxylin and eosin.
Hematoxylin colors nuclei blue Eosin colors the cytoplasm pink
in order to check out the morphology of the tissue and to determine that what you are looking for is present in the section
(e.g., cytoplasm, intra- and extra-cellular filaments, etc.)
• Common stains seen in this class use a
combination of basic and acidic dyes:
H&E, PAS, Trichrome
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Hematoxylin & Eosin (H&E)
Hematoxylin only Eosin only Both
Basic dye: stains only Acidic dye: stains only
Combined, H&E nuclei, ribosomes, and cytoplasm and other provides contrast among other chemically acidic chemically basic cell organelles substances substances
It is used to demonstrate: • glycogen in cell • mucus in certain cells • the basement membrane that underlies epithelia • reticular fibers in connective tissue
Trichrome Stains • Trichrome = “Three colors” • Usually consists of an acidic dye, a basic dye, and a dye for a specific structure or cell product (e.g. lipids, elastic fibers, etc.)
Weigert’s Elastic Stain Masson’s Trichrome Stain
Integumentary system stained with different trichrome stains
Use of images for educational purposes only
Seeing in 2D, Thinking in 3D
Sectional macro- and micro-anatomy allows us to study the topographical relationships among structures
human head human abdomen sagittal slices , human skull C: 3D reconstruction of face Use of images for educational purposes only
21 1/6/2020
In Histology, We have to Reconstruct the 3D Image in our Minds
SEM image of a renal corpuscle
& surrounding kidney tubules Things to keep in mind: • The size and internal structural appearance are reflected in the plane of section • Understanding the gross structure of the organ will help you to interpret your sections • This takes a lot of practice, but is a skill you will use A LOT in professional school! Use of images for educational purposes only
Relative Sizes of Eukaryotic
and Prokaryotic Cells
44
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Scale
Use of images for educational purposes only
Review of Linear Measurements in the Metric System
