Module 2.1 Flashcards
How to convert micrometres to nanometres
multiply *1000
the early microscopes vs modern ligh microscopes
-they both have an eyepiece lens , hey both have a stage for the speciment .
-both have a focsing dial .
-could mgnifya round 300 times .
-Wheareas a MODERN microscope can mgnify round 1000 times .
adavantges of light microscopes
-can be used on living cells , so we can explore porcesses such s cell division or movement of cells .
-cheap and easy to use
-portable .
disadvantages of light microscopes .
-need to use a stain to do this , and these can kill cells .
-Limtied resolution due to the nature of light itself .
-low mgnificationsm ( cannot see smaller strucutres)
A-level definition of resolution .
-Resolution is defined as the minium distance between two objetcs where they can sitll be seen as two separae objects .
What do you mean as the NATURE OF LIGHT .
-For a light microscope , the limit of resolution is around 200nm .
-The wavelength of visible light is around 400nm-700nm , if two objects are closer than 200 nm we cannot see tjem s two separate objetcs using a light microscopes.
Better to use an elecron microscope .
length of wavelength of light microscope
400-700 nm
How does a light microscope work ?
-light passes through the sample , if we were to view certain prt of the cell , the light we see will have already have passed thorugh parts of the cell , making the image blurred nd the resolution worse thn it coudl be .
so what has dramatically improved light microscopy ?
laser scanning confocal microscope .
How does a laser confocal microscope work ?
-A laser s used to scan the object we’re interested in .
-This allows us to view a precise layer f the object .
-Meaning the image is prodced t HIGHER RESOLUTION than conventional light microscopy-observe lviign specimens
Advantages of laser scanning confocal microscopes .
-We can produce three dimensional images of cells .
-HIH MAGNIFICAITON AND RESOLUTION
-This technique allows us to tag specific proteins + srucutures within cells .
-By using speical dyes adn anitbiodies , we can actually see porteins movign around in cells , helpign us to workout the fucntions of those portiens .
Working out magnification ?
Key; always keep all the units the same .
two types of questions on magnificationc an be asked .
-given a scale bar .
-given the equation for magnfiication .
check sheet for quesiton ; how do workout the diemeter of nuclesus (1)
-start by mesuring the length of the scale , abr in millimeers (using a ruelr) . E.g ,t he scal ebr is 10mm .
-READ ACCRUATELY UR RULER .
how do workout the diemeter of nuclesus (2)
Now use a ruler to measure the diameter of the nucleus in mm = 63mm .
workingss
scale bar 10mm
nucleus 63mm
63/10 =6.3 this tells us he nucelus is 6.3x bigger than the scale bar .
how do workout the diemeter of nuclesus (3)
Now we need to workout , how man times bigger the nucleus is compred to the scale br .
how do workout the diemeter of nuclesus (4)
the scale bar represents the distnce of micromentre . do 6.3*1 to get the actual diameter of the nucleus wichh is 6.3 um .
how do you workout out magnicaiiton .
magnificiation = image size / object sixze .
do the question on the sheet .
…
wavelength of laser scanning (confocal)
SAME AS LIGHT MICROSCOPE OR PTICAL micoroscope same thign (400-700 nm)
disadvanatges of laser scanning
-expensive
-requires training
The resolution of any light microscope ,w ill be limited DUE TO THE WAVELENTH OF LIGHT .
So eelctron microscopes were invented .
Why is the resoltuon of n electron light microscope soo good
-Electrons ahvea verys hort wavelength . So the resoltuiton is 2000 times better than a light microscope .
How can particles liek electrons have a wavelength ?
-As electrons have propeorties of both particles and waves sp can have a wavelength .
How does an electron microscope work s1
–Start with an electron gun , which is producing a beam of electrons .
-These electrons pass down the microsocope .
How does an electron microscope work s2
The insidie of an electron microsocpe contains a vacuum .
-So the elctrons can pss through withouth boucning off the molecules in the air .
How does an electron microscope work s3
-As electrons are negaitvelyc hagred , we can focus the electron beam , using electroamgnets . These re clled electromagnetic lenses .
How does an electron microscope work s4
The speciment is placed in both of the electron beams .
-Electrons can pass through some parts of the specimen more easily than other parts .
How does an electron microscope work s5
final image porduced ona fluorescent screen .
Advanatges of electron microscopes .
-Resoloution 2000x better than a light microscope .
-under goood conditiosn ,c an reosolve up to 0.1nm .
-we cana chicve a far greter level of magnificationbefore the image is blurred .
-Ribosomes +structure of cell membrane —> electron microscope .
Wha is the the wavelength odf an electron microscope
short - 0.004nm
Disadavantages of an electron microscope
-Interior is a vacuum ,s oc anot view libign specimens usin an electron microscope .
-Requires very careful staining of the specimen + specimen often has to be veryt hin .
-Can get ARTEFCTS - false imges creted by the staining process or the ocniditons inside the electron microscope . (So bioligusthere to chck what they seeing is real , not an artefct).
2 TYPES OF ELECTRON MICORSCOPE
-trnsmission electron microscopes
-scannign electron microscopes .
aDVANATGES OF TRANSMISSION ELECTRON MICROSCOPES
-ELECTRON BEAM PASSES THORUGH THE SPECIMEN .
-Produces glat 2 dimensional images .
-Very high resolution .
-highest magnifiction
-cleater smaller structures
disadvanatges of transmission electron micorscopes
-onyl works when specimen is thinly sliced .
-non-livign (vacuum)
-large equipment
-lots of training
VERY EXPENSIVE
-b+wvery expensive
How does a transmission electron microscope work ?
-beam of electrons pass thorgh the specimen stained with metal salts .
IT ALSO HAS THE HIGHEST MAGNIFICATIOn out of all microscopes .
SCANNING ELECTRON MICROSCOPE
-the electron beam does not pass through the specimen .
-Instead ,e elctrons are scattered from the surface of he specimen +dteected .
-produce three dimensional images , dpes not require speicmen to be thinlhys lcied.
diadanatge of scannign electorn micorscope
-Thet transmission electron microcope
-lessthan reslutio .
-ALSO requires h speicmen is coated ,w ith ametal sucha s gold , reading toa rtefats .
-very expnesive
-large eequpiment
-lots of training
-non-living cacuum
Slides can be observed the followign ways ;
…
Differentia staining
Dufferentua staining is when you stain a specimen with different stains that bind to specific cell strucutres so that they can be identified within a single prepration .
Stain - methylene blue
all purpose - nucleus dark blue
acetuc orcein
DNA (chromosomes dark red)
Sudan red
lipids red
iodine (potassu=ium idodide solution )
cellulose 9yellow) starch granules (blue /black)
check the sheet use of an eyepiece graticule and stage micrometer
-Mkae sure you are able to convert between all the units REVISE AND LEARN IT .
Why is a stage micrometer used ?
-Stage micrometer allows ut to calibrate the eyepeice graticule so we know how much eye peice graticule actually is .
Callibrate something (1)
-Look at the specimen using eye piecie graticules .
-EXAMPLE —> with of the cell is eyepiece;s (then callibrate this to find out what one eye piece is . )
EXAM TIP , WIDTH OF CELL IS 20 EYEPEICES THEN Cllibrate usign stage micrometer (slide you put on )
Callibrate something (2)
-Focus your microscope under the same magnifiction you looked at your specimen with .
-Make sure your eyepiec egraticule and stage micrometer are PAarallel .
for exmaple , lets say our eye peice uinits is 35 eyepeice units in 1mm
35 eyepiece units = 1mm
1 eyepiece =0.0285mm
1eyepiece=28.5 micrometres
MAKE SURE TO CONVERT INTO MICROMETRES .
THT 20 BEFROM BEFORE 20*28.5 MICROMETRES TO FIND THE WIDTH OF THE CELL .
0.571 MICROMETRES .
CHCK FOR UNITS and decimal places in question /
Exam questions example for callibration
the preparation + examinaiton of microscope slides for use in light micorsocopy .
-nclduign the use of an eyepice graticule and stage micrometr . HAVE aago at the exam questions in ms partners sleeve aswl as PMT .
euakryotic cells
Dna is contained in a membrane bound nuculeus .
(membrane bound –> surrounded by a membrane.)
-some eukaryotic cells can lose their nucleus as the develop (red blood cells i humans .)
-But vast majoirty of eukarytoic cells have a nculeus htorughou their lifespan.
Second key feature of eukarytoic cells
-Dna is tightly wrapped around proteins called histones .
(togehther the DNA AND HISTONE PROTEINS FORM CHROMSOOMES )
What is so good about tightly coling their dna into chromomsomes ?
-euakryotic cells can pack a gret deal of dna into their nucleus .
third key feeatures of eukaryotic cells
-Their Dna is a LINEAR MOLECUL
-“Linear “ , means that the ends of the DNA molecule ina cheomsome ,a re not joined together to form a loop ..
WE ALREADY SEEN - eukarytoic cells have a membrane bound nucleus ….
BUT THEY also contrian nuber of memrbane bound organelles .
All the organelles thar are emmebrane bound in a ANIMAL eukarytoic cells .
-Golgi apparatus .
-Endoplasmic reticulum .
-Mitochocndira .
-cytoplasm
-Nucleus .
All the organelles that are membrane bound in a Eukarytoic cell (Plants)
Everything in animal BUT ALSO
-Vacuole
-Cholroplasts
Fourth key feature of eukaryoitc cells .
-eukaryoitic cells also contain other organelles that are NOT membrane bound .
-AKAA RIBOSOMES .
Ribosomes are invovled in porteins ntheiss .
-Ribosomes are lso in prokaryotic cells
-but the ribosomes in euakryotic cells are larger thant hsoe in prokarytoes .
-Eukarytoic cells conaisns 8OS
The letter S
is simply a unit showinghow quicly organells make in a cneitrfurge
-Eukarytoic cells are surroudned bya cell surface memebrane .
This helps to control the meolcuels that can apss in + out of cells .
BUT , in plants + fngithe clel memebrne by a cell wall .
-Cell wall helps to maintain a strucutre of these cell .
-In plant cells the cell walls s amde of the plyscharide Cellulsoe .
Wht is the cell wall in fungi
polsacharide chitin .
-Animal cells don’t have cell wall .
key differences in prokarytoic cells and eukarytoic cells (1)
-prokaryoic cells are mcuh SMALLWE than eukyartoic cels .
key differences in prokarytoic cells and eukarytoic cells (2)
Prokarytoic cells have NO MEMRABNE MOUND ORAGNELLES AT ALL .
-Therefore , the Dna in prokarytoic cells is in the CYTOPLASMS rther than in the nucelus .
-Unlike eukrytic DNA is arranged into a circular chromosome with NO EFREE ENDS .
-The DNA in prokaryotes is not BOUND to histone proteins .
-Sometimes , bacterial cells contain small loops of DNA called plsmids .
-Plasminds usually contain a reltively small number of GENES .
-But these cn incldue genes which make the bacterium RESISTANT , to antibiotics .
-So plasmids are very important for bacterial .
CCONTRSTS EUAKRYOTES WHIVH DO NOT CONTAIN PLASMIDS .
PROKARYOTIC CELLS ALSO HAVE RIBOSOMES FOR Protein synthesis .
(but hty are smaller than eukarytoic cells )
-prokaryotic cells hav size 70s .
Prokarytoic cells (like bacteria) are surroudned y a cell wall .
-In bacteria , this is made from pentidoglcyen . Which is called murein .
-Peptidoglycen is a polymer formed between peptieds and polysccharide moelcuels .
-he bacterial cell wall helps to maintain the strcuture of the cell . (e.g water moves into the bacterial cell by osmosis , cell wall prevents the cell from bursing .)
Some bacteria porduce a slime cpsule ont he outside of the cell wll .
-slime capsules can help to rpotect the bacteira from PHAGOCYTISIS by white blood cells .
Other strucures of prokaryotic cells (1)
-Proakryotic cells ahve FLAGELUM –> helps them to move
- sperm (eukarytoic) also has flagellum ,
-But prokarytoic cells flagellum DIF trcutrr to eukayrotic flagellum
key differences in prokarytoic cells and eukarytoic cells (2)
-Some surfaces also have fine portein strans on their surfce clled pili .
-Pili help bacteria to attach to surfaces + also to attch to toher bacteira .
-When two bacteria are attached DNA cn be TRANSFERRED from one bacterium to another .
key differences in prokarytoic cells and eukarytoic cells (3)
-Lstly bacteria also contain lipid droplets + glycogen granuls .
-They act s nucirent stores for bacterial cells .
-Sometimes using an electron microscope ,w e can see infoldings in the cell memebrne of prokaryote cells .
-These are clled MESOSOME .
-Initially scientists thoguht they played a role in respiration .
-NOW scienists believe mesossomes are actuallh RTEFCTS . That are creted when bacterial cells re prepared for electron microscopy .