bio101 narvaez
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DIFFERENT TYPES OF MICROSCOPYBY JAN ROBIN D. NARVAEZ
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TYPES OF MICROSCOPY
Fluorescence microscop
!"e #rom Microscop u.com
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TYPES OF MICROSCOPY
Con#oc!l microscop
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TYPES OF MICROSCOPY
Mul$ip%o$on microscop
Im!"e #rom Ol mpusMicroscop Resource
Cen$er &e'si$e
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F()ORESCENCE MICROSCOPY
Fluorescence is $%e m!in componen$
I$ is $%e li"%$ pro+uce+ ' ! su's$!nce*%en i$ is s$imul!$e+ ' !no$%er li"%$
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F()ORESCENCE MICROSCOPY
2o* i$ *or3s4
Cellul!r componen$s !re colorless !n+c!nno$ 'e +is$in"uis%e+ un+er !microscope
Fluorescence microscop s$!inscomponen$s *i$% + es
)V is use+ $o loc!$e cer$!in componen$s
o# ! cell
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F()ORESCENCE MICROSCOPY
2o* i$ *or3s4Im!"e pro+uce+ is '!se+ on $%e secon+
li"%$ source
Im!"e #rom ScienceE+uc!$ion Resource
Cen$er 5 C!rle$on Colle"e
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F()ORESCENCE MICROSCOPY
Principles o# 6uorescence microscop 4Ener" is !'sor'e+ ' $%e !$om1
*%ic% 'ecomes e7ci$e+ T%e elec$ron 8umps $o ! %i"%erener" le,el
T%e elec$ron +rops '!c3 $o $%e
"roun+ s$!$e1 emi$$in" ! p%o$on
Im!"e #rom $%e o9ci!l*e'si$e o# $%e No'el Pri:e
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F()ORESCENCE MICROSCOPY
Fluoroc%romes; 6uorescen$ s$!ins
Arci+ine or!n"e; use+ in +e$ec$ion o#m!l!ri!
Aur!mine; use+ in +e$ec$ion o#$u'erculosis
Fluorescen$ !n$i'o+ use+ in+e$ec$ion o# speci?c !n$i'o+
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F()ORESCENCE MICROSCOPY
Aur!mine;r%o+!mine process
)ses ! ello* 6uorescen$ + e $o ,isu!li:eMycobacterium tuberculosis
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F()ORESCENCE MICROSCOPY
Aur!mine;r%o+!mine s$ep;' ;s$epprocess4
@. Re!"en$s nee+e+ #or s$!inin"
Aur!mine;p%enol solu$ion
Aci+;!lco%ol #or +ecolori:!$ion
Po$!ssium perm!n"!n!$e or !cri+ineor!n"e #or coun$ers$!inin"
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F()ORESCENCE MICROSCOPY
Aur!mine;r%o+!mine s$ep;' ;s$epprocess con$inue+
-. E7!mine un+er $%e 6uorescen$microscope usin" ! lo* po*er o'8ec$i,e
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F()ORESCENCE MICROSCOPY
Aur!mine;r%o+!mine s$ep;' ;s$epprocess con$inue+
. P!$ien$ $es$s posi$i,e #or $u'erculosis*i$% $%e minimum o# #our AFB in $%een$ire sme!r
&%ile ne"!$i,e *%en $%ere is less $%!n#our '!cilli
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F()ORESCENCE MICROSCOPY
A+,!n$!"es42!s $%e !'ili$ $o isol!$e in+i,i+u!l
pro$einsDi eren$ molecules c!n 'e s$!ine+ *i$%+i eren$ colors
Dis!+,!n$!"es4No$ 'ein" !'le $o e7!mine specimens in$%eir li,e or n!$ur!l sel#
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CONFOCA( MICROSCOPY
2o* i$ *or3s4
)ses op$ic!l im!"in" $ec%ni ue $oincre!se op$ic!l resolu$ion o# ! micro"r!p%
)ses sp!$i!l ?l$erin" $ec%ni ues $oelimin!$eou$;o#;#ocus li"%$ in specimens
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CONFOCA( MICROSCOPY
2o* i$ *or3s4A pin%ole is pl!ce in #ron$ o#
$%e illumin!$ion source $o !llo*$r!nsmission onl $%rou"% ! sm!ll !re!
(i"%$ source o# ,er %i"% in$ensi$ li3e
l!ser li"%$ is use+(i"%$ re6ec$s o ! +ic%oric mirror1*%ic% +irec$s i$ $o sc!nnin" mirrors
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CONFOCA( MICROSCOPY
C!n process ;D im!"es
Im!"e #roml mpus6uo,ie*.com
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CONFOCA( MICROSCOPY
)se+ !s ! nonin,!si,e im!"in" $ec%ni ue$%!$ %!s pro,e+ use#ul #or in ,i,o
re!l;$ime c $omorp%olo"ic!l !n!l sis o#!n!l sis o# '!sc!l cell c!rcinom! cellsin?l$r!$in" $%e epi+ermis
)ses ! l!ser source
&!$er immersion o'8ec$i,e lens *!s use+!n+ !$$!c%e+ $o $%e s3inCon#oc!l im!"in" cri$eri! #or BCC *!suse+
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CONFOCA( MICROSCOPY
A+,!n$!"es4
Illumin!$es en$ire ,ie* o# ?el+ uni#orml
Does re uire $o process specimen
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CONFOCA( MICROSCOPY
Dis!+,!n$!"es4
2!rm#ul n!$ure o# %i"%;in$ensi$irr!+i!$ion $o li,in" cells !n+ $issues
2i"% cos$ o# purc%!sin" !n+ oper!$in"
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M)(TIP2OTON MICROSCOPY
Pioneere+ ' &in#rie+ Den3Com'ine+ $%e i+e! o# $*o;p%o$on
!'sorp$ion *i$% $%e use o# ! l!ser sc!nner
Im!"e #rom M!7 Pl!nc3Ins$i$u$e &e'si$e
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M)(TIP2OTON MICROSCOPY
2o* i$ *or3s4Mul$iple !'sorp$ion #rom mul$iple p%o$ons
in+uce ! molecul!r e7ci$!$ion o# !m!"ni$u+e e ui,!len$ $o $%e sum o# $%e!'sor'e+ p%o$on ener"iesA$ ,er %i"% p%o$on +ensi$ies1 i$ 'ecomes#or $*o or more p%o$ons $o 'esimul$!neousl !'sor'e+E!c% mul$ipl !'sorp$ion in+uces !molecul!r e7ci$!$ion o# ! m!"ni$u+ee ui,!len$ $o
$%e sum o# $%e !'sor'e+ p%o$on ener"ies
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M)(TIP2OTON MICROSCOPY
J!'lons3i Ener" Di!"r!m
Im!"e #rom Cornell)ni,ersi$ &e'si$e
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M)(TIP2OTON MICROSCOPY
S3in c!ncer +i!"nosis !n+ moni$orin"
I+en$i?c!$ion o# !'norm!ll loc!$e+mel!noc $es
;D im!"in" $ec%ni ue is use+
Fluorescence *!s !lso inclu+e+ in $%e$ec%ni ue
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M)(TIP2OTON MICROSCOPY
A+,!n$!"es4
Pro,i+es ;D im!"es
P%o$o+!m!"e or p%o$o'le!c%in" is "re!$lre+uce+ +ue $o +ecre!se+ e7ci$!$ion ,olume
E7ci$!$ion s p%o$on ener" is in,erselpropor$ion!l $o *!,elen"$%
(on"er *!,elen$% li"%$ m! 'e use+
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M)(TIP2OTON MICROSCOPY
P%o$ople!c%in" r!$es in mul$ipl s$!ine+specimens
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