Hematoxylin and Eosin (H & E)

-       Hematoxylin: (blue color), basic dye that is positively charged ( has OH group). Very likely to stain the nucleus (nucleus is negatively charged---due to negatively charged DNA)

-       Eosin (counterstain): (red Color), acidic dye that is negatively charged (has high tendency for donating its H’s). Likely to stain cytoskeletal structures such as plasma membrane.

Verification of staining result: enzyme digestion

1.     Periodic acid-Schiff (PAS) reaction: PAS stains carbohydrate rich macromolecules (ex. Glycogen in liver). Staining should disappear upon the addition of amylase or diastase (enzyme that digest carbo-rich molecules)>> prove the validity of staining.

2.     Feulgen reation: similar to PAS. But stain DNA and use DNAse to verify. 

it is cleaved by B-gal

can be useful as a reported gene by putting x-gal in with a gene of interest

Atomic Force Microscopy:

1.     relatively new technique. It is used to look at live cell at electron microscopic resolution. Machine feels or scans the surface of the target object using tapping mode or contact mode to gather information. Computers generates pictures based on info gathered.

Confocal scanning microscopy

1.     optical dissection and reconstruction. Similar to CAT scan. Different wavelengths of light used produce various laminations (different color). Main advantages include: (can visualized physiological changes, can produce 3D replication, this technique illuminate one point in cell and do not excite other tissues that it penetrates (thus causing no disruption)). 

1.     Electron microscopy: 

Scanning electron microscope (SEM): blast heavy metal over things we want to visualized, and then use electron screening to detect object. This technique is used to visualize surface structure. This technique must be done in a vacuum.

simple squamous lining the thoracic wall, heart, abdominal body cavities

this MESOderm

what is Serous membrane

consists of a mesothelium plus supporting connective tissue (with a basal lamina between the two).  It contains no glands and lines the peritoneal, pleural and pericardial cavities.

Junction types.  What are the differences

   1)     Zonulaoccludens 

    2)     Zonulaadherens

    3)     Macula adherens

 1)     Zonula occludens  - tight junctions between the lateral sides of cells, but they only exist in small areas and have selectivity as to their junction strength.  they are near the apical side of the cell

    2)     Zonula adherens - larger zones of tight junction that take up a lot of surface

    3)     Macula adherens - aka desmosomes.  point adhesions that can be anywhere on lateral side and add a lot of strength between neighboring cells

stereocilia are all the same height, are extra long (have candle flame like appearance)

they can be found in the epididymis, and hair cells of the ear

metaplasia is the changing of a cell type appearance, perhaps to a more differentiated state.  it is usually reversible.

dysplasia refers to a progressive abnormal appearance of a cell type

tissue that is constantly turning over and proliferating (ie. bone marrow and GI mucosa)

the other types are stable (arrested at G0) or permanent (do not proliferate)

increased number of cells (hyperplasia) in the lactiferous ducts of the breasts of a 28 yo woman is likely to be physiological because she is young and her breasts can grow.

if this was seen in a 70 yo woman who has already gone through menopause, this would be a pathologic finding

basophilia of chromatin fades (the blue color fades) bc DNA is lost

it is lost because of degradation of endonucleases

nucleus shrinking down to nothing, become tight, solid basophilic mass

when this pykonotic nucleus undergoes fragmentation, it's called karyorrhexis

architecture of dead tissues remains, but they are anucleated

this is because enzymes have degraded proteins, namely proteolysase, and there is nothing to clear the cells.  eventually, neutrophils will get them out of the tissue

a local area of this is known as an INFARCT

cheesy necrosis.

often seen in TB, lysed cells are inclosed in distinct inflammatory borders marked by granulomas

in the pancreas bc lipases are activated.

saponification occurs because cleaved fatty acids combine with calicum to form a visible soapy mess

define:

rubor

calor

tumor

dolor

dunctio laesa

redness

warmth

swelling

pain (prostgladins responsible for this)

loss of function

it is the collection fluid in the interstiteum that is not in hydrostatic and oncotic equilibrium.  it is acellular with low specific gravity

edema

the release of histamines (stored in mast cells, causes permeability of venule) and NO (produced by endothelial cells), increased hydrostatic pressure, transudate

the next stage is increased vascular permeability and exudate can happen

decrease flow velocity, decreased fluid, increased RBCs (as well as leukocytes)

blood cells will tumble and stack (rouleaux) and aggregate internally while WBS will be displaced peripherally near the site of injury

leukocytes are slowed down (told to put their blinkers on) by selectins on endothelial cells near site of injury (expression is stimulated by TNF and IL-1 which are cytokines and are produced by activated macrophages)

adhesion is possible when TNF/IL-1 start expressing integrins  the neutrophil's native integrins get transformed into their high affinity state

margration between the cells is stimulated by chemokines which also come from neutrophils that have already passed through the cell layer and a circular cell-cell messaging cascade is created

 a negative effect Pharmaceuitcals can have on margination process?

opsinization, opsinins are attached to cell surface so they can be identified by other cells (neutrophils have membrane receptors for these cells)

phagocytosis, if oxygen dependent, uses myeloperoxidase, if not, uses enzymes

-chronic granulomatous disease is absence of NADPH oxidase so foreign materials can be engulfed, but never destroyed

serious= transudate fluid, effusion, blisters

fibrinous= increased cellularity in fluid, mainly fibrin

suppurative (purulent)= exudative, pus (which is a bunch of dead WBCs, microbes, and debris), abscess

neutrophils

macrophages and leukocytes and plasma cells

promotes scarring

potent fibrogenic agent which simulates fibroblast chemotaxis

branching of preexisting vessels at site of injury or by recruiting progenitor cells

NO promotes vasodialation while ****VEGF***** promotes vessel permeability so that endothelial cells can migrate to the point of action

there is granulation of the tisssue

fibroblasts arrive to provide collagen formation

angiogenesis is taking place

whole process takes days to 3 weeks

why is TGF-beta the most important fibrogenic agent: 

it's a malignant cancer of connective tissue affecting skeletal muscle.

an osteosarcoma affects actual bone