INTRODUCTION TO CELLS,TISSUES, AND MICROSCOPY

Part 2

VIBS 243Larry Johnson

OBJECTIVES

1. PREVIEW CELLULAR ULTRASTRUCTURE2. PREVIEW CELLS, TISSUES, AND ORGANS3. OVERVIEW OF LIGHT AND ELECTRON MICROSCOPY4. PREPARATION OF SPECIMENS - TYPES OF VISIONS5. ULTRASTRUCTURAL FEATURES OF THE CELL AND

ORGANELLES

ORGAN – Two or more types of tissues; larger functional unite.g., skin, kidney, intestine, blood vessels

Introduction to HISTOLOGY

TISSUE – Groups of cells with same general function and texture (texture = tissue)e.g., muscle, nerve

CELL – Smallest unit of protoplasmSimplest animals consist of a single cell.

ORGAN SYSTEM - Several organse.g., respiratory, digestive, reproductive systems

CELL

TISSUE

ORGAN

SYSTEM

PROTOPLASM – Living Substance

cell

tissue

organ system

organ

FOUR BASIC TYPES OF TISSUES IN THE BODY

-----------------------------------------------EPITHELIUM CONNECTIVE TISSUE

MUSCULAR TISSUE NERVOUS TISSUE

• FUNCTIONS: COVER ORGANS, LINE VISCERA AND BLOOD VESSELS, SECRETORY CELLS OF GLANDS

Epithelium

• FUNCTION: THE HISTOLOGICAL GLUE WHICH BINDS THE OTHER TISSUES TOGETHER TO FORM ORGANS, SPECIALIZATIONS INCLUDE BLOOD, CARTILAGE, AND BONE.

CONNECTIVE TISSUE

BLOOD CELLS(CLASSIFIED AS CONNECTIVE TISSUE)

RED CELLS carry oxygen and carbon dioxidebiconcave disks

WHITE CELLS defense against invadersgranules and lobed or indented nuclei

PLATELETS blood clottinganucleate

Gunther von Hagens’Body Worlds

FatMuscle

• FUNCTION: GENERATION OF CONTRACTILE FORCE

MUSCLE

Gunther von Hagens’Body Worlds

Smooth muscle

NERVOUS TISSUE

NERVOUS TISSUEFUNCTIONS: SPECIALIZED FOR THE

TRANSMISSION, RECEPTION, AND INTEGRATION OF ELECTRICAL

IMPULSES

Gunther von Hagens’Body Worlds

EPITHELIUM CONNECTIVE TISSUE MUSCULAR TISSUE NERVOUS TISSUE

Where are these basic tissues located?

Epithelium

Connective tissue

EPITHELIUM CONNECTIVE TISSUE MUSCULAR TISSUE NERVOUS TISSUE

Where are these basic tissues located?

EPITHELIUM CONNECTIVE TISSUE MUSCULAR TISSUE NERVOUS TISSUE

Where are these basic tissues located?

Muscular tissue

EPITHELIUM CONNECTIVE TISSUE MUSCULAR TISSUE NERVOUS TISSUE

Where are these basic tissues located?

NERVOUS TISSUE

Let’s enjoy some images192 Eyehttp://viewer.serenusview.com/Viewer.aspx?SlideId=54223cf4-223f-4367-85f7-f611d735feef

242 Esophagus and trachea, monkeyhttp://viewer.serenusview.com/Viewer.aspx?SlideId=ae5864df-d2ae-4fc4-814b-8e5e9956c39f

220 Fetal fingerhttp://viewer.serenusview.com/Viewer.aspx?SlideId=ec150f61-6449-43c0-8a4b-a07c110d2573

http://viewer.serenusview.com/Viewer.aspx?SlideId=54223cf4-223f-4367-85f7-f611d735feefhttp://viewer.serenusview.com/Viewer.aspx?SlideId=54223cf4-223f-4367-85f7-f611d735feefhttp://viewer.serenusview.com/Viewer.aspx?SlideId=ae5864df-d2ae-4fc4-814b-8e5e9956c39fhttp://viewer.serenusview.com/Viewer.aspx?SlideId=ae5864df-d2ae-4fc4-814b-8e5e9956c39fhttp://viewer.serenusview.com/Viewer.aspx?SlideId=ec150f61-6449-43c0-8a4b-a07c110d2573http://viewer.serenusview.com/Viewer.aspx?SlideId=ec150f61-6449-43c0-8a4b-a07c110d2573

• FUNCTIONS: RESPONSIBLE FOR THE “IMMUNE RESPONSE” TO FOREIGN INVADERS WHICH IS MEDIATED BY EITHER ANTIBODY PRODUCED BY THE CELLS OR BY THE CELLS THEMSELVES

LYMPHOID TISSUE

BLOOD VESSELSnot one of the four tissues

HISTOLOGICAL IDENTIFICATION:All vessels are lined with endothelium

ARTERY – THICK WALL COMPOSED OF SMOOTH MUSCLE PLUS SOME CONNECTIVE TISSUE CAPILLARY –NARROW TUBE LINED WITH A SINGLE ENDOTHELIAL CELL

VEIN – LARGE LUMEN RELATIVE TO THICKNESS OF CONNECTIVE TISSUE AND SMOOTH MUSCLE WALL

LYMPHATIC – SMALL THIN WALLED VESSELS WHICH CARRY LYMPH

196 Human spermatic cord• http://viewer.serenusview.com/Viewer.aspx?SlideId=652c6183-a5df-4790-b78f-e3f56eef3145

http://viewer.serenusview.com/Viewer.aspx?SlideId=8f70a9e4-ad7b-4542-830d-11beb6596ebf

126 Vein and bile duct

272 Human uterushttp://viewer.serenusview.com/Viewer.aspx?SlideId=a4e2a33a-6005-4831-aeb7-9e85dc8e204e

http://viewer.serenusview.com/Viewer.aspx?SlideId=652c6183-a5df-4790-b78f-e3f56eef3145http://viewer.serenusview.com/Viewer.aspx?SlideId=652c6183-a5df-4790-b78f-e3f56eef3145http://viewer.serenusview.com/Viewer.aspx?SlideId=8f70a9e4-ad7b-4542-830d-11beb6596ebfhttp://viewer.serenusview.com/Viewer.aspx?SlideId=8f70a9e4-ad7b-4542-830d-11beb6596ebfhttp://viewer.serenusview.com/Viewer.aspx?SlideId=a4e2a33a-6005-4831-aeb7-9e85dc8e204ehttp://viewer.serenusview.com/Viewer.aspx?SlideId=a4e2a33a-6005-4831-aeb7-9e85dc8e204e

MAGNIFICATION VS. RESOLUTION

1. MAGNIFICATION - INCREASE IN IMAGE SIZE2. RESOLUTION - SMALLEST DISTANCE BETWEEN TWO

POINTS THAT CAN BE SEEN (DISTINGUISHED)

CALCULATED BY0.61 (WAVELENGTH)/NUMERICAL APERTURE

0.25 um FOR LIGHT MICROSCOPE0.1 nm FOR ELECTRON MICROSCOPE

SAMPLE PREPARATION1. FIXATION2. EMBEDDING

A. PARAFFINB. PLASTIC

3. SECTIONINGA. 0.5 um FOR LIGHT MICROSCOPYB. 60-80 NM FOR ELECTRON MICROSCOPY

STAINING1. LIGHT MICROSCOPY

A. HEMATOXYLIN AND EOSIN (H&E)

B. PERIODIC ACID/SHIFF (PAS)C. TOLUIDINE BLUE

2. ELECTRON MICROSCOPY (TEM)

A. OSMIUMB. LEAD CITRATE

STAINING1. LIGHT MICROSCOPY

A. HEMATOXYLIN AND EOSIN (H&E)

B. PERIODIC ACID/SHIFF (PAS)

C. TOLUIDINE BLUE

Color provides clues

ShapeSizeIntensity of staining

STAINING1. LIGHT MICROSCOPY

A. HEMATOXYLIN AND EOSIN (H&E)

B. PERIODIC ACID/SHIFF (PAS)C. TOLUIDINE BLUE

2. ELECTRON MICROSCOPY (TEM)

A. OSMIUMB. LEAD CITRATE

Fundicstomach (H&E)

145

http://viewer.serenusview.com/Viewer.aspx?SlideId=152b1321-36f5-4695-bd76-f378ee9351d8

Fundic stomach, monkey (PAS)

243

http://viewer.serenusview.com/Viewer.aspx?SlideId=3d651332-f20b-4710-a72b-1d2df28d1087

Surface mucus cells of Fundic stomach, rabbit (toluidine blue)

244

http://viewer.serenusview.com/Viewer.aspx?SlideId=8f512434-a356-49b9-bed1-01d508a10451

BRIGHT FIELD

PHASE CONTRAST

NOMARSKIdifferential interference contrast

DARK FIELD

Dead stained cells

Live unstained cells

OTHER LIGHT AND ELECTRON MICROSCOPE PROCEDURES

• IMMUNOFLUORESCENCE• AUTORADIOGRAPHY• IN SITU HYBRIDIZATION• FREEZE FRACTURE

– (MEMBRANE ANALYSIS)

AUTORADIOGRAPHYself radioactive

Typical TEM

Carbon replica TEM

• Cells in cork• walled boxes that are similar to tiny

rooms, or cellula, occupied by monks = "cell.“

Cellsin a plant

How Did Cells Get Its Name?

Cellin an animal

Cell Size

Cell

LightMicroscopy

ElectronMicroscopy

Cells Contain Organelles

Organelles in cellsare like organs

in animal/human bodies

Cells Contain Organelles

Cell membrane

Cells Contain Organelles

Cell membrane

Marks limit of cells/environment

Marks limit of cells/environment

Cells Contain Organelles

NucleusArchive of cell’s DNA

Archive of cell’s DNA

Cells Contain Organelles

Mitochondria

Produces ATP (cell energy)

Produces ATP (cell energy)

Cells Contain Organelles

Endoplasmic ReticulumProduces proteins and hormonesand detoxifies

Produces proteins and hormonesand detoxifies

Cells Contain Organelles

Golgi ApparatusModifies (adds sugar to)and packages proteins

Modifies (adds sugar to)and packages proteins

Animal Cell

Plant Cell

internetsource

internetsource

https://www.google.com/search?q=images+of+animal+cells&rlz=1C1CHFX_enUS549US549&espv=2&biw=1253&bih=864&source=lnms&tbm=isch&sa=X&ei=7M9oVMzWMoKryASEoYDQCg&ved=0CAYQ_AUoAQ&dpr=0.75#facrc=_&imgdii=_&imgrc=nrxsX86Ul3uqDM%253A%3B0AA4-OVGz6_DEM%3Bhttp%253A%252F%252Fwww.animalcute.net%252Fwp-content%252Fuploads%252F2012%252F04%252FAnimal-Vs-Plant-Cell1.jpg%3Bhttp%253A%252F%252Ftoplowridersites.com%252Fplant-cells-vs-animal-cells-plants-and-animals-are%252F%3B704%3B438

http://www.hart.k12.ca.us/lamesa/staff/teams/8th/brainiacs/jpaul/images/animalCalive.jpg

• Double layer of phospholipids• controls the flow of

water• marks outer limit of cell• separates cell from

environment

Animal Cell

Plant Cell

Animal Cell

Plant Cell

• Has a double membrane• Holds DNA

• Involved in cell division• Involved in directing protein

production by ribosomes

Animal Cell Plant Cell

Eukaryotic Cell

eukaryotic cellhas a nucleus

Animal Cell

Plant Cell

• Make energy for the cell• Can be different shapes• Has a double membrane

Animal CellPlant Cell

Has a double membrane

• Membrane bound sac • Intracellular digestion• Release of cellular waste• Generally small in animal cells

Animal Cell

Small Vacuoles

Electron MicroscopeImage of a Pancreatic Cell

Animal Cell

Animal Cell

• Part of cytoskeleton of the cell• Ring of nine groups of fused microtubules• Groups of three microtubules• Involved in cell division

• Plants do not have centrioles

Centriole

Electron MicroscopeImage of a Animal Cell

Animal Cell

• Contain enzymes necessary for intracellular digestion• In white blood cells, these lysozymes digest bacteria• Cause cell death if improperly released into cytoplasm

Animal Cell

Electron MicroscopeImage of a Nerve

Lysosome

Animal Cell

Electron MicroscopeImage of a Nerve

Lysosome

Animal Cell

CELL COMPOSITION

THREE MAJOR CLASSES OFCYTOPLASMIC STRUCTURE

--------------------------------------------------------------------------------1. MEMBRANOUS ORGANELLES - COMMON STRUCTURES,

METABOLIC FUNCTION, CELL MEMBRANE, RER, SER, GOLGI, MITOCHONDRIA, LYSOSOMES

2. NON-MEMBRANOUS ORGANELLES – CYTOSKELETAL COMPONENTS, MICROTUBULES, MICROFILAMENTS, INTERMEDIATE FILAMENTS, FREE RIBOSOMES

3. INCLUSIONS- EXPENDABLESa. NUTRIENT e.g., GLYCOGEN, LIPIDb. PIGMENT e.g., MELANIN GRANULESc. SECRETORY GRANULE e.g., ZYMOGEN GRANULE OF PANCREAS

In summary

Castle Rock Big Bend National Park TX

The end of

INTRODUCTION TO CELLS,�TISSUES, AND MICROSCOPY �Part 2�OBJECTIVES�Introduction to HISTOLOGYSlide Number 4FOUR BASIC TYPES OF TISSUES IN THE BODY �-----------------------------------------------Slide Number 6CONNECTIVE TISSUEBLOOD CELLS�(CLASSIFIED AS CONNECTIVE TISSUE)�Slide Number 9Slide Number 10MUSCLESlide Number 12NERVOUS TISSUESlide Number 14Slide Number 15Slide Number 16Slide Number 17Slide Number 18Let’s enjoy some imagesLYMPHOID TISSUEBLOOD VESSELS�not one of the four tissues 196 Human spermatic cordSlide Number 23MAGNIFICATION VS. RESOLUTIONSAMPLE PREPARATIONSTAINING STAINING STAINING Fundic �stomach (H&E)Fundic stomach, monkey (PAS)�Surface mucus cells of Fundic stomach, rabbit (toluidine blue)Slide Number 32Slide Number 33Slide Number 34OTHER LIGHT AND ELECTRON MICROSCOPE PROCEDURES Slide Number 36Slide Number 37Slide Number 38Slide Number 39Slide Number 40Cells Contain OrganellesCells Contain OrganellesCells Contain OrganellesCells Contain OrganellesCells Contain OrganellesCells Contain OrganellesCells Contain OrganellesAnimal CellSlide Number 49Slide Number 50Slide Number 51Slide Number 52Slide Number 53Slide Number 54Slide Number 55Slide Number 56Slide Number 57Slide Number 58Slide Number 59Slide Number 60CELL COMPOSITION In summarySlide Number 63The end of