HistoLec- Monthly

Human Histology with Cytogenetic Histology- study of tissues - Microscopic anatomy Cytogenetic- study of chromosomes 12 Steps of Tissue Processing 1. Numbering - Also known as logging/recording - Process of assigning the number to a specimen - 3 important parts: 1.Sequence of reception - 004 = fourth specimen received 2.Indicate if S or A - S stands for Surgical meaning the tissue came from the OR (buhay pa yung source) - A stands for Autopsy meaning the tissue came from the morgue (patay na yung source) 3.Year the is specimen received - Record all information on a logbook indicating the specimen source 2. Fixation - Process of preservation - Main goal: Preserve the tissue in a life-like manner - Stop autolysis/ putrefaction - Routine fixative: 10% formalin 3. Dehydration - Process of removing water from the tissues - It can be achieved by increasing grades of Alcohol(ethanol) to avoid tissue distortion - We start at 70%- 80% - 90%- 95%- Absolute alcohol (99.99%, not 100%), but for delicate tissues like brains and embryos, we start at 50%. - 2.5 mm x3 mm x 0.5 mm is the size of the tissue from the organ to be processed. 4. Clearing - Ultimate goal is to make tissues clear or transparent - It also removes the excess alcohol - Routine clearing agent: Xylene/ Xylol 5. Paraffin infiltration - Also called as paraffin impregnation - We immerse the tissue in melted paraffin wax - The melted paraffin wax will fill-up the tissue cavities
Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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HistoLec- Monthly
6. Embedding - Aka as casting/blocking - Process of placing the tissue in a precise position inside a mold. - Most common mold is the paper boat - Melted paraffin wax is poured on the paper boat until is filled. This will serve as paraffin bed. - Using forceps, the tissue will be placed in the middle of the paraffin bed. - Melted paraffin wax will be poured again to cover of the paper boat. Wait until it solidifies. - End result of embedding is paraffin bloc or tissue block. 7. Trimming - Trim-off the excess paraffin wax - The surrounding paraffin wax should only be 2mm. - Shape the tissue block as a truncated pyramid (Smaller upper portion than basal portion.) 8. Sectioning - Aka Microtomy (To make use of a microtome) - The process of cutting the tissues into thin slices called sections or ribbons with a thickness of 4-6 9. Staining - Minor steps before staining: 1. Subject the tissue in a floatation water bath. - For flattening the wrinkles and crumples - The water bath has a black background, 11 inches in diameter and 4 inches in height. Pour 2 litres of water. Add tissue adhesive. 2.Fish-out the tissues. - We make use of glass slides and then immerse it and chase the ribbons. 3.Deparaffinization - We are removing the paraffin wax by melting it. - Four ways of removing paraffin wax: 1.Bunsen burner 2.Slide blower 3.Incubator 4.Paraffin Oven (most common; we set it 2 to 5 degrees higher that the materials melting point. Usually 60 degrees for paraffin) - Process of promoting optical differentiation - We use H and E staining technique. - H means Hematoxylin which is a basic dye that stains acidic structures like the nucleus. It gives the color black to blue. It is alsao called as the nuclear stain.
Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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HistoLec- Monthly
E means Eosin which is an acidic dye that stains basic structures like the mitochondria. It gives the color reddish to pink to orange. It is also called as the cytoplasmic stain.

10. Mounting - It is the process of cementing slides and coverslips - Its main goal is to preserve the stain. - We use mounting medium like Eukit and Canada Balsam 11. Labelling - We indicate the use of special stains *Decalcification- optional process between Fixation and Dehydration Process of removing calcium from the specimen Applicable if teeth, bones, teratomas (mass of tissue which when cut will exhibit epithelial outgrowth) Also known as demineralization Decalcifying agent: Mild Acid like 10% HNO3 Cell Structure and Function Cell Theory Rudolf Virchow All living things are made up of cells. Cells are the smallest working units of all living things. All cells come from preexisting cells through cell division. Definition of Cell A cell is the smallest unit that is capable of performing life functions. Examples of Cells Two Types of Cells Prokaryotic Do not have structures surrounded by membranes Few internal structures One-celled organisms, Bacteria Eukaryotic Contain organelles surrounded by membranes Most living organisms Typical Animal Cell

Three Major Parts of a Cell PROTOPLASM living substance within a cell that sustains life

Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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HistoLec- Monthly
1.Cytoplasm - It is the living space around the nucleus and it is where organelles are located. 2. Karyoplasm - Inside the Nucleus or Nucleoplasm CYTOSKELETON For the maintenance of the cell INCLUSIONS Deposits of fats, carbohydrates and proteins that are not necessarily needed to maintain life of the cell.

Cell Parts Surrounding the Cell Cell Membrane Also called Plasmalemma or Plasma membrane COMPOSITION: Phospholipids Cholesterol - To reduce the fluidity of the cell membrane Oligosaccharides (glycocalyx) - Cell coat that allow cell to cell interaction CHONS (Proteins) For structural support, maintenance and dimensional shape 1. Integral Proteins - These are the transmembrane proteins that are integrated in phospholipid bilayer One-pass partially embedded (1 layer of phospholipid bilayer) Two-pass fully embedded 2. Peripheral Proteins - Underneath the phospholipid bilayer FUNCTIONS: 1. Selective barrier 2. Recognition 3. Cell-to-cell interaction 4. Structural integrity 5. Acts as interface between the cytoplasm & external environment MOLECULAR STRUCTURE Nicholson and Singer Fluid-mosaic model 2 Leaflets: Phospholipids CHONS
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1:1 ang ratio pero kapag myelinated, the phospholipids outweigh the CHON 4:1

Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

HistoLec- Monthly
Inside the Cell Nucleus Directs cell activities Separated from cytoplasm by nuclear membrane Contains genetic material DNA Nuclear Membrane Surrounds nucleus Made of two layers Openings allow material to enter and leave nucleus Made-up of pores Chromosomes In nucleus Made of DNA Contain instructions for traits & characteristics Nucleolus Inside nucleus Contains RNA to build proteins Cytoplasm Gel-like mixture Surrounded by cell membrane Contains hereditary material Endoplasmic Reticulum Largest membranous system With vesicles and tubules FUNCTIONS: 1. CHON synthesis and modification 2. Lipid and steroid synthesis 3. Detoxification of certain toxic compounds 4. Manufacture of membrane of the cell Smooth type: lacks ribosomes Rough type (pictured): ribosomes embedded in surface Smooth ER More tubular than RER Has become specialized in some cells sarcoplasmic reticulum FXN: synthesis of phospholipid membrane and steroids Rough ER More flattened Prominent in cells specialized for CHON secretion Myelin sheath can be seen in nervous tissues Unit membrane: trilaminar structure

Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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HistoLec- Monthly
Ribosomes Protein synthesis 4 segments of rRNA Intensely basophilic 40 s and 60s Mitochondria Powerhouse of the cell Produce ATP/Energy Self-replicate due to cDNA Specialized for aerobic respiration & production of ATPs highly plastic L.M. : eosinophilic E.M.: 2 compartments Outer membrane - sieve-like - porins Inner membrane - Folded - Cardiolipin - cristae Contact sites Intermembrane space Intercristal space Golgi Bodies Protein modification, packaging, sorting Move materials within the cell Move materials out of the cell 3 levels of cisternae 1. cis-face *closest to RER *convex *entry face 2. medial face *trans-face *concave *exit face Lysosome Digestive 'plant' for proteins, fats, and carbohydrates Transports undigested material to cell membrane for removal Cell breaks down if lysosome explodes pH 5.0

Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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HistoLec- Monthly
PEROXISOMES Self-replicating microbodies Spherical to ovoid Enzymes: 1. Urate oxidase 2. Catalase 3. D-amino acid oxidase Fxn: catabolism of long-chained fatty acids PROTEASOMES FXN: ubiquitination (non-functional peptide) Morphology: -cylindrical structure -made up of 4-stacked rings; each w/ 7 CHONS

Let them give thanks to the LORD for His unfailing love and His wonderful deeds for men. - Psalms 107:8

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