Treść książki

Przejdź do opcji czytnikaPrzejdź do nawigacjiPrzejdź do informacjiPrzejdź do stopki
Oneofthemostimportantandubiquitousspectroscopictechniquewhich
hasemergedinthemostlabsisfluorescencespectroscopy,thistechniquesis
usedbyscientistsofmanydisciplinesfromatomicphysicstotheallfieldsof
LifeSciences.Bytheway,herealsoitisworthtonotethatfluorescenceisone
oftheoldestscientificmethodwhichservesfromtheearlierbeginningof
modernphysicsera.Letusremindthattheseriesofhydrogenatomspectral
lineswerediscoveredduetofluorescenceofH-atoms,thisinformationwas
crucialandservedasaninitialpointforcreatingmodernquantumphysics.
Later,duringafewdozensyearsfluorescencewassuccessfullyexploredin
studydifferentmolecularsystemsinvariousmaterials,semiconductors,
plasmasofdifferentkinds,spacematter,solids,organicandbiomolecules
andmorecomplicatedorganicobjectsfromcellstobiologicaltissues.
Duringthepast20yearsattheendof20thcenturytherehasbeenremark-
ablegrowthintheuseoffluorescencespectroscopyintheLifeSciences,i.e
inbiology,medicineandinthedevelopmentofnewtechnicalmethodsand
apparatusforresearches.Conventionalandtimeresolvedfluorescencespec-
troscopyareconsideredtobeprimaryresearchtoolsinbiochemistryandbi-
ophysics.Thisemphasishaschangedduringthelastdecade,andtheuseof
fluorescencehasexpandeddramatically.Fluorescenceemissionexcitedby
differentsourcesfromconventionalsourcesoflighttoshotpulsesoflasers
andnanoseconddischargesingasesisnowadominantspectroscopicmeth-
odologyusedextensivelyinbiotechnology,flowcytometry,medicaldiag-
nostics,DNAsequencing,forensics,andgeneticanalysis,wenamefew.The
thingisthatfluorescencedetectionishighlysensitive,andthereisnolonger
theneedfortheexpenseanddifficultiesofhandlingradioactiveisotopesfor
mostbiochemicalmeasurements(Lakowicz2006).Therehasbeendramatic
growthintheuseoffluorescenceforcellularandmolecularimaging,stand-
ardapparatusnowareavailablefromleadingworldmanufacturingcompa-
niessuchasOlympus,KarlZeiss,Hitachiandanumberofothers.Fluores-
cenceimagingcanrevealthelocalizationandmeasurementsofintracellular
molecules,sometimesatthelevelofsinglemolecules.
Obviouslyonecanputthequestion,whatdistinguishesfluorescencespectrosco-
pyfromalltheothermethodssuggestedforstudyingprocesseswithcomplexcom-
poundsparticipation?Primarilyitisitsultra-highsensitivityaswasmentioned
videsupra.Obviously,thisfeatureisespeciallydemandedwhenthe
fluorophorexistsintraceamounts.Withproperdyeselectionandproper
experimentalconditionstheabsolutesensitivitymayreachthelimitofsingle
moleculeswhichisrealizedinconventionalconfocallasermicroscopyar-
rangement.Thisissufficientandveryattractiveformanyapplications,par-
6