Fluorescence microscopy: dzekutanga nzira

Absorção de Água uye apiwa akabuda chiedza mberi inorganic uye Organic zvinwiwa mugumisiro phosphorescence kana fluorescence. Kusiyana mananara ndiyo nguva yacho penguva pakati Absorção de Água uye akabuda chiedza nekubudisa. Kana fluorescence pamusoro azvaiitwa kuitika anenge panguva, apo phosphorescence - nevamwe kunonoka.

nhoroondo mashoko

Muna 1852, musayendisiti British Stokes, akatanga kurondedzera fluorescence. Iye akatanga idzva izwi somugumisiro pakati kuedza pamwe fluorspar, izvo emit chiedza dzvuku pasi ultraviolet chiedza. Stokes akaona nezvechimwe chinhu chinonakidza. Unowanikwa kuti wavelength ari fluorescent mwaranzi anogara kupfuura unyatsoshanda excitation chiedza.

Kuti asimbise pfungwa muzana remakore rechi19 kwave kuedza zhinji. Vakaratidza kuti siyana ivhu fluoresce pasi pokudzora ultraviolet chiedza. Pakati zvinhu, pakati pezvimwe zvinhu, zvave crystals, Inogadzirwa nemasiridzi anobuda mumiti, zvicherwa, chlorophyll, ichakasangana nezvimwewo zvinodhaka, inorganic yeimwe, mavhitaminzi, mafuta. Kushandiswa madhayi nokuti miedzo zvoutachiona zvakananga akatanga chete muna 1930

Fluorescence microscopy: tsananguro

Vamwe zvakashandiswa mune yokutanga yezana zvidzidzo rechi20 remakore akaratidza kumusoro specificity. стал важнейшим инструментом и в биомедицинских, и в биологических исследованиях. Nemhaka kuita, aisagona kuitwa zvakasiyana nzira, iyo nzira fluorescence microscopy rava chinhu chinokosha kushandisa anorapa achishandisa mitemo uye chaivo tsvakurudzo. Zvakafanana zvabuda zvinokosha avapo, uye nokuda zvokuvakisa.

? Zvakanakirei sei nzira fluorescence microscopy? Kushandisa zvinhu zvitsva wava zvichibvira uye kusarudzwa zvikuru chaizvo muchitokisi submicroscopic zvinoriumba. Fluorescence microscopy anogona kuona mamorekuru kuroorwa. A-siyana madhayi kubvumira kuziva vakawanda zvinhu pakarepo. Pasinei shoma spatial kupedza pamusoro diffraction panogumira midziyo, izvo zvinozoita kunobva zvakananga ehupfumi dzokuenzanisira, ari kuziva mamorekuru pazasi ichi pamwero kuri ingangodaro. Various mimwe pashure irradiation zviratidzwa autofluorescence. Fani iyi zvinowanzoshandiswa petrology, bhotani, ari semiconductor indasitiri.

Features

Dzidza munyama mhuka kana utachiona kazhinji akaoma kana simba kana simba kwazvo unspecific autofluorescence. Zvisinei, kukosha muzvidzidzo anowana sumo kupinda zvinhu zvinoumba inovapo pane imwe wavelength uye emitting anodiwa chiedza chekubuda zvikuru. Fluorochromes vave madhayi vaikwanisa vakazvimiririra kurerekera zvivako (zvinoonekwa kana asingaoneki). Saka vane matunhu selectivity kuti kubaya, uye quantum goho.

стала широко применяться с появлением естественных и синтетических красителей. Fluorescence microscopy rave richishandiswa nevakawanda kubva veMubatanidzwa dzinongoitika uye chakaita zvokugadzirwa madhayi. Zvavainazvo vamwe kusimba Profiles pamusoro anobuda uye excitation uye nangananga kuti yakananga utachiona kushungurudza.

Kuziva munhu mamorekuru

Kazhinji, mune akanaka ezvinhu, unogona kunyora imwe yoga maatomu akafanana Naiwo mufaro. Nokuti ichi, pakati pezvimwe zvinhu, zvakakodzera kupa zvakakwana pasi mheremhere detector uye Manyiminya background. Fluorescein Morekuru kuti kukundikana nokuda photobleaching kunogona emit photons kusvika zviuru. 300. At 20% muunganidzwa kunyatsoshanda acho uye anogona kuvanyora mune mari inenge 60 zviuru.

, основанная на лавинных фотодиодах или электронном умножении, позволяла исследователям наблюдать поведение отдельных молекул на протяжении секунд, а в ряде случаев и минут. Fluorescence microscopy kwakavakirwa aval photodiodes kana yemagetsi chokuwanziridza, akabvumira vaongorori kuchengeta maitiro nomunhu mamorekuru kuburikidza wechipiri, uye mune zvimwe zviitiko kunyange maminitsi.

kuoma

Chinhu chikuru nyaya vachida kudzvinyirira pakati Manyiminya ruzha background. Nokuti vazhinji zvakashandiswa pamasikiro filters uye lenses kuratidza zvimwe autofluorescence, zvinoitwa masayendisiti mukutanga mumatanho vakanga nezvatinovavarira pagadzirwe yenhengo kuva pasi fluorescence. Zvisinei, zvakazoitika kuedza zvakakonzera mhedziso itsva. , основанная на полном внутреннем отражении, позволяет достичь низкого фона и высокоинтенсивного возбуждающего светового потока. Zvikurukuru, kwakaonekwa kuti fluorescence microscopy, yakavakirwa zvachose womukati yokufungisisa, hunobvumira kuti vabudirire pasi yaipinda uye yakakwirira zvikuru excitation chiedza.

mashandiro

, основанной на полном внутреннем отражении, заключаются в использовании быстрозатухающей или нераспространяющейся волны. Misimboti fluorescence microscopy, yakavakirwa zvachose womukati kuratidzwa kuri kushandiswa evanescent chinozunguzirwa kana evanescent. It ndepana muganhu pakati vezvenhau nedzimwe refractive indices. Panyaya iyi, chiedza danda inopfuura nomunyika prism. It ane yakakwirira refractive indekisi parameter.

The prism yakabatana aqueous mhinduro kana girazi pamwe yakaderera parameter. Kana danda chiedza kutaurirwa pana necheparutivi kuti zvikuru dzinonetsa, danda kunooneka zvachose inowanikwa. Fani Izviwo, kunokonzera nonpropagating mafungu. Nemamwe mashoko, wakadenha remagetsi munda chinopinda kupinda svikiro ane nechepazasi refractive indekisi parameter kune kure asingasviki 200 nanometers.

The evanescent chinozunguzirwa kusimba chiedza zvaizova zvakakwana kuti fadza fluorophores. Zvisinei, nemhaka ayo udzamu zvikuru diki, vhoriyamu chayo achava duku. Zvinoguma nechekumashure yakaderera-chechetere.

nadzurudzo

Fluorescence microscopy pazvikanganiso zvachose womukati yokufungisisa, unogona chinozoiswa pamwe epi-jekeso. Izvi zvinoda lenses dzakakwirira kusanganiswa aperture (kanenge 1,4, asi chinhu chinodiwa kuti yakasvika 1.45-1.6), uye zvishoma zvakavhenekerwa munda midziyo. The yokupedzisira kunokwanisika duku gwapa saizi. Nokuti mukuru kufanana uchishandisa nhete mhete, izvo nemutemo chikamu yorwizi. Nokuti dzinonetsa yorusvingo, pashure iyo pane zvachose yokufungisisa, tinofanira yakakwirira azvipire refraction vanobhabhatidzwa svikiro iri izvo nechifukidziro Girazi maikorosikopu.