Fluorescence Microscopy: ụkpụrụ nke usoro

Absorption na re-emission nke ìhè n'ihu inorganic na organic oke mmiri bụ n'ihi phosphorescence ma ọ bụ fluorescence. The dị iche n'etiti phenomena bụ oge nke n'agbata absorption na emission nke ìhè agbanwe oge. Mgbe fluorescence ndị a Filiks ime ihe fọrọ nke nta n'otu oge, mgbe phosphorescence - na ụfọdụ oge.

ya mere eme

Na 1852, British ọkà mmụta sayensị Stokes, mbụ kọwara fluorescence. Ọ ẹkenam a ọhụrụ okwu dị ka a n'ihi nke nwere na fluorspar, nke emit red ìhè n'okpuru ultraviolet ìhè. Stokes kwuru ihe na-akpali onu. Ọ na-achọta na wavelength nke fluorescent radieshon bụ mgbe ukwuu karịa eruba nke excitation ìhè.

Iji gosi na amụma na narị afọ nke 19 na e nwere ọtụtụ nwere. Ha gosiri na a dịgasị iche iche nke samples fluoresce n'okpuru nduzi nke ultraviolet ìhè. N'etiti ihe, ihe ndị ọzọ, a na kristal, resins, minerals, chlorophyll, crude ọgwụ ọjọọ, inorganic ogige, vitamin, mmanụ. The kpọmkwem ojiji nke e ji esiji ákwà maka ndu ule malitere naanị na 1930

Fluorescence Microscopy: nkọwa

Ụfọdụ n'ime ihe ndị na-eji ke akpa ọkara narị afọ nke 20 ọmụmụ gosipụtawo elu specificity. стал важнейшим инструментом и в биомедицинских, и в биологических исследованиях. Ekele arụmọrụ, nke ike-enweta site na iche na ụzọ, na usoro nke fluorescence Microscopy aghọwo ihe dị oké mkpa na ngwá ọrụ na Biomedical na ndu nnyocha. Dịkwa mkpa pụta nọ na-enwetara, na maka ihe.

? Gịnị uru ka usoro nke fluorescence Microscopy? Iji ihe ọhụrụ ndị na-ekwe omume na nhọrọ nke ukwuu kpọmkwem cell submicroscopic mmiri. Fluorescence Microscopy nwere ike ịchọpụta otu ụmụ irighiri. A dịgasị iche iche nke e ji esiji ákwà ekwe njirimara nke multiple ihe ozugbo. N'agbanyeghị ole gbasara ohere mkpebi nke diffraction ịgba nke bụ ngwá ọrụ, nke, n'aka, na-adabere na kpọmkwem Njirimara nke sample, njirimara nke ụmụ irighiri n'okpuru a n'ogo bụkwa ezi omume. Dị iche iche n'omume mgbe irradiation ngosi autofluorescence. Nke a na onu na ọtụtụ-eji petrology, botani, na okporo ọkụ ụlọ ọrụ.

atụmatụ

Omumu anụmanụ anụ ahụ ma ọ bụ pathogens mgbe mgbagwoju anya ma ọ bụ na ike na-adịghị ma ọ bụ nnọọ ike unspecific autofluorescence. Otú ọ dị, uru dị na ọmụmụ enweta iwebata n'ime ihe mmiri ụtọ na a kpọmkwem wavelength na emitting ndị dị mkpa ìhè eruba osisi ike. Fluorochromes eme ka e ji esiji ákwà ike nke onwe ha mmasị owuwu (anya ma ọ bụ na-adịghị ahụ anya). N'ihi ya, ha nwere a elu selectivity lekwasịrị, na kwantum mkpụrụ.

стала широко применяться с появлением естественных и синтетических красителей. Fluorescence Microscopy e n'ọtụtụ ebe na ebe ọ bụ na biakwa obibia nke eke na sịntetik dyes. Ha nwere ụfọdụ osisi ike profaịlụ nke emission na excitation na ezubere iche kpọmkwem ndu zaa.

Identification nke onye ụmụ irighiri

Ọtụtụ mgbe, na ọnọdụ ka mma, ị nwere ike ịdenye a iche iche mmewere na-egbukepụ egbukepụ. N'ihi na nke a, ihe ndị ọzọ, ọ dị mkpa iji nye a zuru ezu ala mkpọtụ nke detector na ngwa anya ndabere. Fluorescein molekul na odida ruru photobleaching nwere ike emit ruo 300 puku. Photons. Na 20% collection arụmọrụ nke usoro na pụrụ aha ha na-ego nke banyere 60 puku.

, основанная на лавинных фотодиодах или электронном умножении, позволяла исследователям наблюдать поведение отдельных молекул на протяжении секунд, а в ряде случаев и минут. Fluorescence Microscopy dabeere na nnukwu photodiodes ma ọ bụ electronic multiplication, kwere ka na-eme nnyocha na-edebe omume nke onye ụmụ irighiri site nke abụọ, na ụfọdụ ọbụna nkeji.

complexity

A isi nke na ihu ọma nke suppression nke ngwa anya mkpọtụ ndabere. N'ihi na eziokwu na ọtụtụ n'ime ihe ndị na-eji na imewe nke nzacha na anya m igosipụta ụfọdụ autofluorescence, mgbalị ndị ọkà mmụta sayensị na-amalite amalite na-gbakwasara na mmepụta nke mmiri na-enwe a ala fluorescence. Otú ọ dị, ụdi nwere emewo ka ọhụrụ nkwubi okwu. , основанная на полном внутреннем отражении, позволяет достичь низкого фона и высокоинтенсивного возбуждающего светового потока. Karịsịa, ọ hụrụ na fluorescence Microscopy, dabeere na ngụkọta esịtidem echiche, ọ na-enye ohere iji nweta a ala ndabere na akwa siri excitation ìhè.

usoro

, основанной на полном внутреннем отражении, заключаются в использовании быстрозатухающей или нераспространяющейся волны. Ụkpụrụ nke fluorescence Microscopy, dabeere na ngụkọta esịtidem echiche bụ ojiji nke evanescent efegharị efegharị ma ọ bụ evanescent. Ọ pụtara na ókè-ala n'etiti media na dị iche iche Refractive indices. Na nke a, ìhè doo agabiga prism. Ọ nwere nnukwu Refractive index oke.

The prism n'akụkụ ka okenye ihe ngwọta ma ọ bụ iko na ala oke. Ọ bụrụ na a doo nke ìhè na-eduzi na ya na n'akuku nke bụ ihe dị oké egwu, na doo na-kpam kpam gosipụtara si interface. Nke a na onu, n'aka, na-akpata nonpropagating ebili mmiri. Ndị ọzọ okwu, na site akpa ubi nke penetrates n'ime a na-ajụ na a ala Refractive index oke ka a anya-erughị 200 nanometers.

The evanescent efegharị efegharị osisi ike nke ìhè ga-ezu ejikere fluorophores. Otú ọ dị, n'ihi na nke oké obere omimi, ya olu ga-nnọọ obere. N'ihi bụ a ala-larịị ndabere.

mgbanwe

Fluorescence Microscopy dabeere ngụkọta esịtidem echiche, nwere ike na-emejuputa atumatu na epi-ọkụ. Nke a na-achọ anya m na-elu space aperture (dịkarịa ala 1.4, ma ọ bụ na-achọsi ike na ọ ruru 1.45-1.6), na n'omume ìhè ubi ngwa. Nke ikpeazụ a na-enweta na a obere ntụpọ size. N'ihi na ukwuu Ime Otu eji a mkpa mgbanaka, nke a na-egbochi site a òkè nke iyi. N'ihi na oké egwu n'akuku, mgbe nke na e nwere a ngụkọta echiche, anyị mkpa a elu larịị nke refraction nmikpu na-ajụ na oghere na cover iko mikroskopu.