Questions? Comments? boggusrl@email.uc.edu Image Review #1 Labs 1-3 Questions? Comments? boggusrl@email.uc.edu

LAB #1 “THE CELL”

eosinophilic basophilic STAINS H and E Hematoxylin is blue/basophilic Eosin is pink/eosinophilic/acidophilic eosinophilic basophilic

PAS Stain stuff that starts with G: glycocalyx, goblet cells, glycogen, glycoprotein

There are two of them here, each is trilaminar PLASMA MEMBRANE There are two of them here, each is trilaminar Each is 9nm thick – can’t see well with LM unless juxtaposed (needs to be at least 200 nm to see with LM, FYI)

polysomes RIBOSOMES Stain BASOPHILIC (baso = blue) If you see bluish stuff in the cytoplasm that means the ribosomes are workin’ it out there and the cell is making proteins Polysomes = ribosomes on an mRNA string Look like little swirlies polysomes

Golgi Apparatus Cis face and trans face – stuff buds off of the trans face Looks like a smiley face – trans face is on the upper lip and cis face is on the chin Cis face faces nucleus, trans face faces the outside of cell – makes sense when you think about how things are transported

Meet the EM Golgi – look at the budding vesicles! trans face cis face

Don’t forget about the golgi ghost in LM slides!!

Mitochondria Via oxidative phosphorylation, supply ATP for the cell Cristae? Mg and Ca granules? What other organelle do we see here?

ER Made of cisternae – flattened tubes Two types: RER and SER Can they be connected? YES! The difference? RER has ribosomes, SER does not If you see cytoplasmic basophilia in a LM slide, then there is a lot of RER in the cytoplasm. The basophilia is due to the rRNA of the ribsomes that are attached to the RER.

ER cont’d If you are talking about neurons, cytoplasmic basophilia is called “ergastoplasm” and the RER stains with Nissl stain. Thus, the RER is called Nissl substace. Makes sense.

SER Usually rounder than RER and looks like it has more buds on it. Also, no ribosomes. That always helps. SER SECRET: if you think it may be SER or RER or Golgi or you have no clue, and you can’t decide what to put down, look for the mitochondria if you can. If the mitochondria are really round and circular, there is a good chance that it is SER. this is because in cells that are involved in detox functions (and thus have lots of SER), the mito are rounder.

round mito

RER vs. SER

Lysosomes Have a membrane, UNLIKE peroxisomes, which do not Contain acid phosphatases and acid hydrolases to break stuff down – can stain for these enzymes to find the lysosomes

Peroxisomes Very smooth looking except for a huge granule (not in humans though) Contain catalase and oxidase

Microtubules Microfilaments Intermediate Filaments Cytoskeleton Microtubules Microfilaments Intermediate Filaments

Microtubules (24 nm)

Often will be labeled near the membrane

Don’t forget…. They are involved in the mitotic spindle

CENTRIOLES Involved in cell division Can you find it?

One more just for good measure

Microfilaments -- Made of actin -- Contractile – look for them in muscle -- 7nm – smallest of the cytoskeletal components

More microfilaments They also make up the core of microvilli

Look right by the membrane…. Have a very fine appearance

INTERMEDIATE FILAMENTS 10 nm – intermediate in size between microfilaments and microtubules They are made of protein that are tissue specific A main component of desmosomes……

GLYCOGEN PIGMENT/MELANIN LIPID CELLULAR INCLUSIONS GLYCOGEN PIGMENT/MELANIN LIPID

GLYCOGEN Solid and black in EM Reddish in LM, but you will usually get EM if they ask you about glycogen

LM glycogen from lab

LIPID…. My favorite

More lipid…. Also note the round mito….. What do you see with round mito??? SER. One of SER’s functions is fat metabolism, so makes sense that there is lipid there too.

PIGMENT Usually melanin, so if they are trying to show you pigment it will usually be in the skin, in melanocytes, which are found under the keratinocytes.

The nucleus In all its glory

pore Interphase nucleus Has nuclear membrane – 2 membranes juxtaposed with pores in them Note: nuclear membrane made of intermediate filaments pore

Taking a peek into the nucleus Heterochromatin – dark, bundled up chromatin, inactive Euchromatin – light, unwound, ready to go, active eu nucleolus hetero

granular fibrillar Nucleolus 3 things they could label: Perinuclear chromatin – chromatin pressed onto the outside of the nucleolus Fibrillar portion – the darker stuff Granular portion – the lighter stuff granular fibrillar

Prophase Nuclear membrane disrupted Chromatin looks fibrous, stringy It is condensing to become chromosomes

Metaphase Chromosomes are condensed and lined up in the middle, the mitotic spindle is evident The mitotic spindle converges at centrioles – called the “Microtubule Organizing Center” or the centrosome 3 types of microtubules here 1. Astral MT that radiate out like a star – help with orientation 2. Mitotic spindle MT that go across the spindle; can see these ones the best 3. Microtubules that are attached to the chromosomes

Metaphase

Anaphase Chromosomes are separating Can clearly see the spindle and astral microtubules

Telophase Can start to see new cell membrane between the 2 new daughter cells Chromosomes look more separate again – not as distinct as in anaphase

Karyotype This was on our exam and a lot of people forgot what it was called. Do not be that guy.

Quick Quiz What is the main structure, and also the cytoskeletal element that you see here?

Cilia, with a core of microtubules!

Whats the black stuff??

Glycogen!

Whats the white stuff?? (this is tough)

Holes where lipid used to be Holes where lipid used to be. Remember that lipid usually does not come out in sections!

What are the arrows pointing to?

Microtubules in the axon of a dendrite!

What is the dark organelle shown here?

Lysosomes! Look at all the stuff in it!

Whats this??

On the right is a centriole On the left is the Golgi On the right is a centriole

What is found in these cells?

Melanin! The arrow is pointing at the row of melanocytes under all the keratinocytes in the skin

Whats this??

NUCLEAR PORE!

Questions??? Boggusrl@email.uc.edu The End Questions??? Boggusrl@email.uc.edu