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Thermophiles bụ microorganisms na-eme nke ọma na oke okpomọkụ. Ịmụ ha pụrụ inye ihe ọmụma bara uru banyere otú ndụ si eme ka ọnọdụ ndị dị oké njọ kwekọọ. Otú ọ dị, ọ na-esiri ike ịnweta ọnọdụ okpomọkụ dị elu site na iji microscopes anya nkịtị. Atụpụtala ọtụtụ ihe ngwọta emere n'ụlọ dabere na ọkụ eletrik na-eguzogide mpaghara, mana ọ nweghị ngwọta azụmahịa dị mfe. N'ime akwụkwọ a, anyị na-ewebata echiche nke microscale laser kpo oku n'elu oghere microscope iji nye okpomọkụ dị elu maka ọmụmụ ihe thermophile ka ọ na-eme ka ebe onye ọrụ dị nwayọọ. Enwere ike nweta kpo oku Microscale na ike laser na-agafeghị oke site na iji mkpụrụ nanoparticle mkpuchi ọla edo dị ka ihe na-ekpo ọkụ na-ekpo ọkụ na-arụ ọrụ nke ọma. A na-atụle mmetụta enwere ike nke mkpụkọ mmiri microscale, njide cell, na ngagharị thermophoretic centrifugal. E gosipụtara usoro a n'ụdị abụọ: (i) Geobacillus stearothermophilus, nje bacteria na-arụ ọrụ thermophilic nke na-emepụtaghachi na ihe dịka 65 Celsius, nke anyị hụrụ na-eto eto, na-eto eto na igwu mmiri n'okpuru okpomọkụ microscale; (ii) Thiobacillus sp., ihe kacha mma hyperthermophilic archaea. na 80 ° C. Ọrụ a na-emeghe ụzọ maka nleba anya dị mfe ma dị nchebe nke microorganisms thermophilic site na iji ngwaọrụ microscopy ọgbara ọhụrụ na nke dị ọnụ ala.
N'ime ọtụtụ ijeri afọ, ndụ n'ụwa etolitela iji kwekọọ n'ụdị ọnọdụ gburugburu ebe obibi nke a na-ewere mgbe ụfọdụ dị oke egwu n'echiche mmadụ. Karịsịa, ụfọdụ microorganisms thermophilic (bacteria, archaea, fungi) nke a na-akpọ thermophiles na-eme nke ọma na okpomọkụ sitere na 45 ° C ruo 122 ° C1, 2, 3, 4. Thermophiles na-ebi n'ime ebe dị iche iche dị iche iche, dị ka mmiri mmiri hydrothermal vents, mmiri ọkụ. ma ọ bụ mpaghara mgbawa ugwu. Nnyocha ha ewepụtala mmasị dị ukwuu n'ime iri afọ ole na ole gara aga maka opekata mpe ihe abụọ. Nke mbụ, anyị nwere ike ịmụta n'aka ha, dịka ọmụmaatụ, ka thermophiles 5, 6, enzymes 7, 8 na membranes 9 si kwụsie ike na oke okpomọkụ dị otú ahụ, ma ọ bụ otú thermophiles nwere ike isi nagide oke ọkụ ọkụ10. Nke abụọ, ha bụ ihe ndabere maka ọtụtụ ihe dị mkpa ngwa biotechnological1,11,12 dị ka mmanụ ụgbọala mmepụta13,14,15,16, chemical synthesis (dihydro, alcohols, methane, amino acids, wdg)17, biomining18 na thermostable biocatalysts7 ,11, 13. Karịsịa, mmeghachi omume polymerase chain (PCR) 19 a ma ama ugbu a gụnyere enzyme (Taq polymerase) nke dịpụrụ adịpụ na nje bacteria Thermus aquaticus, otu n'ime thermophiles mbụ a chọtara.
Agbanyeghị, ọmụmụ nke thermophiles abụghị ọrụ dị mfe na enweghị ike imezi ya na ụlọ nyocha ọ bụla. Karịsịa, a pụghị ịhụ thermophiles dị ndụ na vitro site na iji microscope ọkụ ọ bụla, ọbụlagodi na ụlọ kpo oku dị na ahịa, nke a na-edekarị maka okpomọkụ dị obere ka 40 Celsius. Kemgbe 1990s, ọ bụ naanị otu nyocha ole na ole ewepụtala onwe ha maka iwebata sistemu microscopy dị elu (HTM). Na 1994 Glukh et al. A tụụrụ ime ụlọ kpo oku / oyi na-adabere na iji cell Peltier na-achịkwa okpomọkụ nke capillaries akụkụ anọ mechiri iji nọgide na-enwe anaerobicity 20. Enwere ike ikpo ọkụ ruo 100 Celsius C na ọnụego 2 Celsius C / s, na-enye ndị na-ede akwụkwọ ohere ịmụ ihe na-akpali akpali nke nje bacteria hyperthermophilic Thermotoga maritima21. Na 1999 Horn et al. E mepụtala ngwaọrụ yiri nke ahụ, ka dabere na iji capillaries na-ekpo ọkụ dabara maka microscopy azụmahịa iji mụọ nkewa / njikọ sel. Mgbe ogologo oge nke enweghị ọrụ ikwu, ịchọ HTM dị irè maliteghachiri na 2012, karịsịa n'ihe gbasara usoro akwụkwọ nke otu Wirth nke jiri ngwaọrụ Horn et al chepụtara. Afọ iri na ise gara aga, a na-amụ motility nke ọnụ ọgụgụ buru ibu nke archaea, gụnyere hyperthermophiles, na okpomọkụ ruo 100 ° C na-eji capillaries na-ekpo ọkụ23,24. Ha mezikwara microscope izizi iji nweta ọkụ ngwa ngwa (ọtụtụ nkeji kama nkeji 35 iji rute ọnọdụ okpomọkụ a tọhapụrụ) wee nweta gradient okpomọkụ kwụ ọtọ karịa 2 cm gafee etiti ahụ. A na-eji ngwaọrụ na-akpụzi ọnọdụ okpomọkụ a (TGFD) iji mụọ mmegharị nke ọtụtụ thermophiles n'ime gradients okpomọkụ n'ebe dị anya dị mkpa 24, 25.
Ịkpọ capillaries mechiri emechi abụghị naanị ụzọ isi lelee thermophiles dị ndụ. Na 2012, Kuwabara et al. A na-eji ụlọ Pyrex a na-atụfu ihe arụrụ n'ụlọ ejiri ihe nrapado na-eguzogide okpomọkụ mechie (Super X2; Cemedine, Japan). A na-etinye ihe nlele ahụ n'efere ọkụ na-ekpo ọkụ na-ere ahịa (Micro Heat Plate, Kitazato Corporation, Japan) nke nwere ike ikpo ọkụ ruo 110 Celsius, mana ọ bụghị na mbụ maka bioimaging. Ndị ode akwụkwọ ahụ hụrụ nkewa nke ọma nke nje bacteria anaerobic thermophilic (Thermosipho globiformans, oge okpukpu abụọ 24 min) na 65 Celsius. Na 2020, Pulshen et al. E gosipụtara ikpo ọkụ nke ọma nke efere ígwè azụmahịa (Attofluor TM, Thermofisher) site na iji ihe ọkụ ọkụ abụọ emebere n'ụlọ: mkpuchi na ogbo (nhazi nke igwe PCR). Njikọ a na-ebute otu okpomọkụ mmiri mmiri ma gbochie evaporation na condensation na ala nke mkpuchi. Ojiji nke O-mgbanaka na-ezere mgbanwe gas na gburugburu ebe obibi. A na-eji HTM a, nke a na-akpọ Sulfoscope, mee ihe onyonyo Sulfolobus acidocaldarius na 75°C27.
Mkpebi a ghọtara nke usoro ndị a niile bụ mmachi maka iji ebumnuche ikuku, imikpu mmanụ ọ bụla na-ekwesịghị ekwesị maka oke okpomọkụ dị otú ahụ yana maka eserese site na> 1-mm n'ụdị transperent. Mkpebi a ghọtara nke usoro ndị a niile bụ mmachi maka iji ebumnuche ikuku, imikpu mmanụ ọ bụla na-ekwesịghị ekwesị maka oke okpomọkụ dị otú ahụ yana maka eserese site na> 1-mm n'ụdị transperent. Общепризнаныm ностатки всех систем было ограничение na иspolzovannы воздушныh объектов, е погружение в масло не подходило для такой высокой температуры и для визуализации через прозрачне 1. Achọpụtara adịghị ike nke usoro ndị a niile bụ njedebe na iji ebumnuche ikuku, ebe ọ bụ na imikpu mmanụ ọ bụla adịghị mma maka oke okpomọkụ dị otú ahụ na maka ịhụ anya site na nlele anya> 1 mm.所有这些系统的一个公认限制是限制使用空气物镜，任何油浸都不认限制是限制使用空气物镜。厚的透明样品成像. Mmachi a ma ama nke sistemu ndị a niile bụ mmachi nke iji enyo nwere ikuku, n'ihi na imikpu mmanụ ọ bụla adịghị mma maka eserese transperent sample> 1 mm oke na oke okpomọkụ dị otú ahụ. Общепризнаныm ностатки всех систем являеся использование воздушныh, ужение в масло непригодно для таких высоких температур и визуализации через прозрачные образцы толщ1. Ihe ndọghachi azụ nke sistemu ndị a niile bụ obere oghere ikuku ikuku, imikpu mmanụ ọ bụla adịghị mma maka oke okpomọkụ na nhụta anya site na nlele anya> 1 mm oke.N'oge na-adịbeghị anya, Charles-Orzag et al welitere mmachi a. 28, bụ onye mepụtara ngwaọrụ nke na-adịghịzi enye okpomọkụ gburugburu usoro nke mmasị, kama n'ime mkpuchi mkpuchi n'onwe ya, kpuchie ya na obere transperent oyi akwa nke resistor mere nke ITO (indium-tin oxide). Enwere ike kpoo mkpuchi ahụ ruo 75 Celsius C site na ịfefe ọkụ eletrik site na oyi akwa transperent. Otú ọ dị, onye na-ede akwụkwọ aghaghịkwa ikpo ọkụ oghere ahụ n'ebumnobi, ma ọ bụghị ihe karịrị 65 Celsius C, ka ọ ghara imebi ya.
Ọrụ ndị a na-egosi na a naghị anabata mmepe nke microscopy ngwa anya dị elu nke ọma, na-achọkarị ngwá ọrụ arụrụ n'ụlọ, a na-enwetakwa ya mgbe ọ na-efu ụgwọ mkpebi nke oghere, nke bụ nnukwu mgbaghara nyere na microorganisms thermophilic adịghị ibu karịa ole na ole. micrometer. Mbelata kpo oku olu bụ isi ihe na-edozi atọ pụta ụwa nsogbu nke HTM: ogbenye oghere mkpebi, elu thermal inertia mgbe usoro okpomọkụ elu, na-emerụ kpo oku nke gburugburu (mmanụ imikpu, ebumnobi oghere ... ma ọ bụ onye ọrụ aka) na oké okpomọkụ. ).
N'ime akwụkwọ a, anyị na-ewebata HTM maka nlele thermophile nke na-adabereghị na ikpo ọkụ. Kama, anyị nwetara kpo oku mpaghara n'ime oke mpaghara nke oghere microscope site na ọkụ ọkụ laser nke mkpụrụ na-amịpụta ọkụ. A na-ahụ anya nkesa okpomọkụ site na iji microscopy quantitative phase (QPM). A na-egosipụta ịdị irè nke usoro a site na Geobacillus stearothermophilus, nje bacteria na-ekpo ọkụ na-ekpo ọkụ nke na-amụpụta ihe dị ka 65 Celsius C ma nwee obere oge okpukpu abụọ (ihe dị ka nkeji 20), na Sulfolobus shibatae, hyperthermophile nke na-eto nke ọma na 80 ° C (archaea). iji maa atụ. A na-ahụ ọnụ ọgụgụ mmụgharị nkịtị na igwu mmiri dị ka ọrụ okpomọkụ. Nke a laser HTM (LA-HTM) ejedebeghị site na ọkpụrụkpụ nke mkpuchi ma ọ bụ site na ọdịdị nke ebumnobi (ikuku ma ọ bụ mmanụ immersion). Nke a na-enye ohere iji oghere ọ bụla dị elu dị na ahịa mee ihe. Ọ naghị ata ahụhụ site na ikpo ọkụ ngwa ngwa n'ihi inertia thermal (na-enweta ọkụ ngwa ngwa n'ọtụtụ millisecond) ma na-eji naanị ngwa ahịa dị. Naanị ihe nchekwa nchekwa ọhụrụ metụtara ọnụnọ nke ọkụ laser dị ike (nke na-abụkarị 100 mW) n'ime ngwaọrụ ahụ yana ikekwe site na anya, nke chọrọ enyo mkpuchi.
Ụkpụrụ nke LA-HTM bụ iji laser kpoo ihe nlele na mpaghara n'ime oghere nke microscope (Fig 1a). Iji mee nke a, ihe nlele ahụ ga-abụrịrị ọkụ. Iji a ezi uche laser ike (ihe na-erughị 100 mW), anyị adabereghị na absorption nke ìhè site na mmiri mmiri na-ajụ, ma artificially ụba absorption nke sample site na mkpuchi nke mkpụrụ na gold nanoparticles (Fig. 1c). Ịkpọ ọkụ nanoparticles ọla edo na ọkụ bụ ihe dị mkpa na mpaghara plasmonics thermal, yana ngwa a na-atụ anya na biomedicine, nanochemistry ma ọ bụ iwe ihe ubi anwụ29,30,31. N'ime afọ ole na ole gara aga, anyị ejirila LA-HTM a n'ọtụtụ ọmụmụ metụtara ngwa plasma thermal na physics, kemistri na bayoloji. Isi ihe siri ike na usoro a bụ igosipụta profaịlụ okpomọkụ ikpeazụ, ebe ọ bụ na okpomọkụ dị elu na-ejedebe na mpaghara microscale n'ime ihe nlele ahụ. Anyị egosila na enwere ike nweta nkewa ọnọdụ okpomọkụ site na iji interferometer transverse shear interferometer nke nwere ogologo ogologo ogologo anọ, usoro dị mfe, nke dị elu na nke nwere mmetụta dị ukwuu nke microscopy nke quantitative nke dabere na iji gratings akụkụ abụọ (nke a makwaara dị ka cross gratings). 33,34,35,36. Ntụkwasị obi nke usoro microscopy a na-ekpo ọkụ, dabere na gafere grating wavefront microscopy (CGM), egosipụtala n'ime akwụkwọ iri na abụọ e bipụtara n'ime afọ iri gara aga37,38,39,40,41,42,43.
Atụmatụ nke echichi nke yiri laser kpo oku, nhazi na okpomọkụ microscope. b Sample geometry nke nwere ọnụ ụlọ AttofluorTM nwere ihe mkpuchi mkpuchi na-acha ọla edo nanoparticles. c Lee anya nke ọma na nlele (ọ bụghị ka ọnụ ọgụgụ). d na-anọchi anya profaịlụ laser doo edo na (e) nkesa okpomọkụ na-esote n'usoro n'ụgbọ elu ihe atụ nke nanoparticles ọla edo. f bụ profaịlụ laser beam nke afọ nke kwesịrị ekwesị maka ịmepụta otu okpomọkụ dị ka egosiri na ịme anwansị nke nkesa okpomọkụ na-apụta na (g). Ogwe ọnụ ọgụgụ: 30µm.
Karịsịa, anyị nwetara kpo oku nke mkpụrụ ndụ anụ mammali na nso nso a na LA-HTM na CGM wee nyochaa nzaghachi okpomọkụ cellular n'ogo nke 37-42 Celsius C, na-egosipụta ntinye nke usoro a na foto cell dị ndụ. Otú ọ dị, ngwa LA-HTM na-amụ banyere microorganisms na okpomọkụ dị elu abụghị ihe mgbagwoju anya, n'ihi na ọ na-achọ ịkpachara anya karị ma e jiri ya tụnyere mkpụrụ ndụ mammalian: nke mbụ, kpo oku ala nke ọkara site na iri iri nke ogo (kama ole na ole) na-eduga. gaa na gradient okpomọkụ kwụ ọtọ siri ike. nwere ike ịmepụta convection nke mmiri 44 nke, ma ọ bụrụ na ejikọtaghị ya na mkpụrụ, nwere ike ime ka mmegharị na-adịghị mma na ngwakọta nke nje bacteria. Enwere ike ikpochapụ convection a site na ibelata ọkpụrụkpụ nke oyi akwa mmiri mmiri. Maka ebumnuche a, na nnwale niile ewepụtara n'okpuru ebe a, a na-etinye nkwusioru nje n'etiti mkpuchi abụọ dịka 15 µm gbara ọkpụrụkpụ etinyere n'ime iko igwe (Attofluor TM, Thermofisher, Fig. 1b, c). Na ụkpụrụ, enwere ike ịzere convection ma ọ bụrụ na ọkpụrụkpụ nke mmiri mmiri dị ntakịrị karịa nha ọkụ nke laser kpo oku. Nke abuo, ịrụ ọrụ na geometry nwere oke dị otú ahụ nwere ike ịkụda ihe ndị dị ndụ aerobic (lee fig. S2). Enwere ike izere nsogbu a site na iji mkpụrụ nke na-adaba na ikuku oxygen (ma ọ bụ gas ọ bụla ọzọ dị mkpa), site n'ịhapụ ikuku ikuku tọrọ n'ime ihe mkpuchi ahụ, ma ọ bụ site n'ịgbapụta oghere na mkpuchi elu (lee Fig. S1) 45 . N'ime ọmụmụ ihe a, anyị họọrọ ngwọta ikpeazụ (Figure 1b na S1). N'ikpeazụ, ikpo ọkụ laser adịghị enye nkesa okpomọkụ otu. Ọbụlagodi n'otu ike nke eriri laser (Fig 1d), nkesa okpomọkụ abụghị otu, kama ọ dị ka nkesa Gaussian n'ihi mgbasa ọkụ (Fig 1e). Mgbe ihe mgbaru ọsọ bụ guzobe kpomkwem okpomọkụ na ubi nke echiche maka ọmụmụ usoro ndu, uneven profaịlụ na-adịghị mma na nwekwara ike iduga thermophoretic ije nke nje bacteria ma ọ bụrụ na ha adịghị agbaso na mkpụrụ (lee Fig. S3, S4)39. Iji mezuo nke a, anyị na-eji a spatial light modulator (SLM) na-akpụzi infrared laser doo dị ka ọdịdị nke mgbanaka (Fig. 1f) na ụgbọ elu nke sample iji nweta a n'ụzọ zuru okè edo okpomọkụ nkesa n'ime a nyere geometric mpaghara. n'agbanyeghị mgbasa ọkụ (Fig 1d) 39, 42, 46. Tinye ihe mkpuchi n'elu n'elu efere ígwè (Njirimara 1b) iji zere evaporation nke ọkara ma lelee ma ọ dịkarịa ala ụbọchị ole na ole. N'ihi na emechighị ihe mkpuchi elu a, enwere ike ịgbakwunye ọkara nke ọzọ ngwa ngwa n'oge ọ bụla ma ọ dị mkpa.
Iji gosi ka LA-HTM si arụ ọrụ ma gosipụta na ọ bara uru na nyocha thermophilic, anyị mụrụ nje bacteria Geobacillus stearothermophilus, nke nwere oke okpomọkụ na-eto eto gburugburu 60-65ºC. Nje bacteria nwekwara flagella na ike igwu mmiri, na-enye ihe ọzọ na-egosi ọrụ cellular nkịtị.
A na-ebute ihe nlele (Fig 1b) na 60 Celsius C maka otu awa wee tinye ya na ihe nrịbama LA-HTM. Nke a tupu incubation bụ nhọrọ, mana ọ ka bara uru, n'ihi ihe abụọ: Nke mbụ, mgbe agbanyere laser, ọ na-eme ka mkpụrụ ndụ na-eto ngwa ngwa ma kewaa (lee ihe nkiri M1 na ihe mgbakwunye). Na-enweghị tupu incubation, nje bacteria na-egbu oge ihe dị ka nkeji 40 oge ọ bụla a na-ekpo ọkụ na ebe nlele ọhụrụ na nlele ahụ. Nke abuo, 1 hour tupu incubation kwalitere adhesion nke nje bacteria na mkpuchi, na-egbochi sel ịpụpụ n'ọhịa nke anya n'ihi thermophoresis mgbe laser agbanyere (lee ihe nkiri M2 na Mmeju Materials). Thermophoresis bụ mmegharị ahụ nke ụmụ irighiri ihe ma ọ bụ ụmụ irighiri ihe n'akụkụ gradient okpomọkụ, na-abụkarị site na ọkụ ruo oyi, na nje bacteria adịghị ewepu43,47. A na-ewepụ mmetụta a na-adịghị mma n'elu mpaghara enyere site na iji SLM na-eme ka ọkpụkpụ laser wee nweta nkesa okpomọkụ dị larịị.
Na fig. Ọnụọgụ 2 na-egosi nkesa okpomọkụ nke CGM na-enweta site na ịgbanye mkpụrụ osisi iko nke ejiri ọla edo nanoparticles nwere eriri laser annular (Fig 1f). A hụrụ nkesa okpomọkụ dị larịị n'akụkụ ebe niile nke ọkụ laser kpuchiri. Edobere mpaghara a ka ọ bụrụ 65ºC, ọnọdụ okpomọkụ kacha mma. N'èzí mpaghara a, usoro okpomọkụ na-adaba na \(1/r\) (ebe \(r\) bụ nhazi radial).
Maapụ okpomọkụ nke nha CGM enwetara site na iji eriri laser annular iji mee ka oyi akwa nke nanoparticles ọla edo na-eme ka ọ nweta profaịlụ okpomọkụ dị larịị n'elu mpaghara okirikiri. b Isotherm nke map okpomọkụ (a). Akara ntụ ntụ na-acha ntụ ntụ na-anọchi anya ihe nhịahụ nke ọkụ laser. Emegharịrị nnwale ahụ ugboro abụọ (lee Ngwa mgbakwunye, eserese S4).
A na-enyocha ike nke mkpụrụ ndụ nje bacteria ruo ọtụtụ awa site na iji LA-HTM. Na fig. 3 na-egosi nkeji oge maka onyonyo anọ ewepụtara na ihe nkiri elekere 3 nke nkeji iri abụọ (Ihe nkiri M3, Ozi mgbakwunye). A hụrụ na nje bacteria na-arụsi ọrụ ike na-agbasa n'ime okirikiri nke laser kọwara ebe okpomọkụ dị mma, na-eru nso 65 Celsius. N'ụzọ dị iche, uto cell na-ebelata nke ukwuu mgbe okpomọkụ dara n'okpuru 50 ° C maka 10 s.
Onyonyo omimi anya nke nje bacteria G. stearothermophilus na-eto mgbe ọkụ ọkụ laser gasịrị n'oge dị iche iche, (a) t = 0 min, (b) 1 h 10 min, (c) 2 h 20 min, (d) 3 h 20 min, nke 200 ewepụtara na ihe nkiri nkeji otu nkeji (ihe nkiri M3 ewepụtara na ozi mgbakwunye) nke etinyere na maapụ okpomọkụ kwekọrọ. Laser na-agbanye n'oge \(t=0\). Agbakwunyela isotherms na onyonyo siri ike.
Iji nwetakwuo quantify cell ibu na ya dabere na okpomọkụ, anyị tụrụ mmụba nke biomass nke dị iche iche chịrị nke mbụ dịpụrụ adịpụ nje bacteria na Movie M3 ubi (Fig. 4). A na-egosi nje bacteria ndị nne na nna ahọpụtara na mmalite nke nhazi otu mini colony (mCFU) na Figure S6. Ejiri igwefoto CGM 48 were were tụọ nha igwe kpọrọ nkụ nke ejiri mee maapụ nkesa okpomọkụ. Ikike nke CGM iji tụọ ịdị arọ na okpomọkụ bụ ike nke LA-HTM. Dị ka a tụrụ anya, okpomọkụ dị elu mere ka nje bacteria na-eto ngwa ngwa (Fig 4a). Dị ka e gosiri na ọkara log ibé na fig. 4b, uto na okpomọkụ niile na-esote exponential uto, ebe data na-eji exponential ọrụ \(m={m}_{0}{10}^{t/\ tau }+ {{ \mbox{cst}}}\), ebe \(\tau {{{{\rm{log }}}}}2\) – oge ọgbọ (ma ọ bụ oge okpukpu abụọ), \( g =1/ \tau\) - ọnụ ọgụgụ uto (ọnụọgụ nkewa kwa nkeji oge). Na fig. 4c na-egosi ọnụọgụ uto dị iche iche na oge ọgbọ dịka ọrụ nke okpomọkụ. A na-eji mCFU na-eto ngwa ngwa site na saturation nke uto mgbe awa abụọ gachara, omume a na-atụ anya ya n'ihi njupụta nje dị elu (dị ka oge kwụ ọtọ na omenala mmiri mmiri oge gboo). Ụdị izugbe \(g'aka ekpe (T' aka nri)\) (Fig 4c) dabara na usoro abụọ a na-atụ anya maka G. stearothermophilus na oke uto dị mma gburugburu 60-65 ° C. Dakọtara data ahụ site na iji ụdị kaadị (Figure S5)49 ebe \(\ekpe({{G}_{0}{;\;T}}_{{\min }};{T}_{{opt}} ; {T}_{{\max}}\right)\) = (0.70 ± 0.2; 40 ± 4; 65 ± 1.6; 67 ± 3) °C, nke kwekọrịtara nke ọma na ụkpụrụ ndị ọzọ e zoro aka na ya na akwụkwọ49. Ọ bụ ezie na a na-enwe ike imepụtagharị paramita dabere na ọnọdụ okpomọkụ, oke uto nke \({G}_{0}\) nwere ike ịdịgasị iche site n'otu nnwale gaa na nke ọzọ (lee ọnụọgụ S7-S9 na ihe nkiri M4). N'ụzọ dị iche na paramita kwesịrị ekwesị nke okpomọkụ, nke kwesịrị ịbụ eluigwe na ala, ọnụ ọgụgụ kachasị elu na-adabere na njirimara nke ọkara (ịnweta nri, ikuku oxygen) n'ime geometry microscale hụrụ.
a Microbial uto na dị iche iche okpomọkụ. mCFU: Miniature Colony Forming Unit. Data enwetara site na vidiyo nke otu nje bacteria na-eto na gradient okpomọkụ (ihe nkiri M3). b Otu dị ka (a), ọnụ ọgụgụ ọkara-logarithmic. c Ọnụego uto (\tau\) na oge ọgbọ (g\) gbakọrọ site na nkwụghachi azụ linear (b). Ogwe mperi mperi: oke okpomọkụ nke mCFU gbasaara n'ọhịa nlele n'oge uto. Ogwe mperi mperi: mperi ọkọlọtọ regression linear.
Na mgbakwunye na uto nkịtị, ụfọdụ nje bacteria na-ese n'elu anya mgbe ụfọdụ n'oge okpomọkụ laser, nke bụ omume a na-atụ anya maka nje bacteria na flagella. Ihe nkiri M5 na ozi ndị ọzọ na-egosi mmemme igwu mmiri dị otú ahụ. N'ime nnwale a, a na-eji ụzarị laser edo edo mepụta gradient okpomọkụ, dịka egosiri na foto 1d, e na S3. Ọgụgụ 5 na-egosi usoro onyonyo abụọ ahọpụtara na ihe nkiri M5 na-egosi na otu nje bacteria na-egosipụta mmegharị ntụzịaka ebe nje bacteria ndị ọzọ na-anọgide na-enweghị mmegharị.
Oghere oge abụọ (a) na (b) na-egosi igwu mmiri nke nje bacteria abụọ dị iche iche nwere okirikiri nwere ntụpọ. Ewepụtara foto ndị a na ihe nkiri M5 (nke e nyere dị ka ihe mgbakwunye).
N'ihe banyere G. stearothermophilus, mmegharị ahụ na-arụ ọrụ nke nje bacteria (Fig 5) malitere sekọnd ole na ole ka agbanyere ọkụ laser. Nlebanya a na-emesi nzaghachi nwa oge nke microorganism thermophilic a na mmụba nke okpomọkụ, dịka Mora et al hụburu. 24 . Enwere ike nyochaa isiokwu nke motility nje bacteria na ọbụna thermotaxis site na iji LA-HTM.
E kwesịghị inwe mgbagwoju anya na igwu mmiri microbial na ụdị mmegharị anụ ahụ ndị ọzọ, ya bụ (i) mmegharị nke Brownian, nke yiri ka ọ bụ ihe mgbagwoju anya na-enweghị ntụzịaka doro anya, (ii) convection 50 na thermophoresis 43, nke na-agụnye mmegharị nke mmegharị mgbe niile na okpomọkụ. gradient.
A maara G. stearothermophilus maka ikike ya ịmepụta spores na-eguzogide nke ukwuu (nhazi spore) mgbe ekpughere ọnọdụ ọjọọ gburugburu ebe obibi dị ka ihe nchebe. Mgbe ọnọdụ gburugburu ebe obibi na-adị mma ọzọ, spores na-etolite, na-etolite mkpụrụ ndụ dị ndụ ma maliteghachi ito eto. Ọ bụ ezie na a maara usoro sporulation / germination nke ọma, ọ dịtụbeghị mgbe a hụrụ ya ozugbo. Iji LA-HTM, anyị na-akọ ebe a nlebanya mbụ nke germination omume na G. stearothermophilus.
Na fig. 6a na-egosi ihe oyiyi oge na-adịghị anya nke omimi anya (OT) nwetara site na iji usoro CGM nke 13 spores. Maka oge nchịkọta dum (15 h 6 min, \ (t = 0 \) - mmalite nke ikpo ọkụ laser), 4 n'ime 13 spores germinated, na oge na-aga n'ihu \ (t = 2 \) h, \ ( 3 \). h \(10 \)', \(9\) h \(40\)' na \(11\) h \(30\)'. Ọ bụ ezie na ọ bụ naanị otu n'ime ihe omume ndị a ka egosiri na Figure 6, a pụrụ ịhụ ihe omume 4 germination na ihe nkiri M6 na ihe mgbakwunye. N'ụzọ na-akpali mmasị, germination yiri ka ọ bụ enweghị usoro: ọ bụghị ihe ọkụkụ niile na-eto eto ma ghara ịmị n'otu oge, n'agbanyeghị mgbanwe ndị ahụ na ọnọdụ gburugburu ebe obibi.
Oge nkwụsị nke nwere ihe oyiyi 8 OT (mikpu mmanụ, 60x, 1.25 NA ebumnobi) na (b) evolushọn biomass nke G. stearothermophilus aggregates. c (b) Edepụtara ya n'ọkara ndekọ ọnụ ọgụgụ ka ọ pụta ìhè n'ahịrị nke ọnụego uto (akara dashed).
Na fig. 6b,c na-egosi biomass nke ọnụ ọgụgụ sel n'ọhịa nlele dị ka ọrụ nke oge n'ime oge niile nke nchịkọta data. Nri ngwa ngwa nke oke akọrọ ahụrụ na \(t=5\) h na fig. 6b, c, n'ihi ọpụpụ nke ụfọdụ mkpụrụ ndụ n'ọhịa nke anya. Ọnụego uto nke mmemme anọ a bụ \(0.77pm 0.1\) h-1. Uru a dị elu karịa ọnụego uto jikọtara ya na Figure 3. 3 na 4, ebe mkpụrụ ndụ na-eto eto. Ihe kpatara mmụba nke mmụba nke G. stearothermophilus sitere na spores adịghị edozi, ma nha ndị a na-egosipụta mmasị nke LA-HTM ma na-arụ ọrụ na otu cell larịị (ma ọ bụ na otu mCFU larịị) ịmụtakwu banyere mgbanwe nke ndụ cell. .
Iji gosipụta ngbanwe nke LA-HTM na arụmọrụ ya na oke okpomọkụ, anyị nyochara uto nke Sulfolobus shibatae, hyperthermophilic acidophilic archaea nwere okpomọkụ kachasị mma nke 80 ° C51. E jiri ya tụnyere G. stearothermophilus, archaea ndị a nwekwara ụdị ọdịdị dị iche iche, nke yiri 1 micron spheres (cocci) karịa elongated rods (bacilli).
Ọgụgụ 7a nwere ihe onyonyo omimi nke usoro S. shibatae mCFU nwetara site na iji CGM (lee ihe nkiri atụmatụ M7 na Ngwa mgbakwunye). MCFU a na-eto na gburugburu 73 Celsius, n'okpuru okpomọkụ kacha mma nke 80 Celsius C, mana n'ime oke okpomọkụ maka uto na-arụ ọrụ. Anyị hụrụ ọtụtụ ihe omume fission mere mCFU dị ka micrograpes nke archaea ka awa ole na ole gachara. Site na onyonyo OT ndị a, a tụrụ mCFU biomass ka oge na-aga wee gosipụta ya na eserese 7b. N'ụzọ na-akpali mmasị, S. shibatae mCFUs gosipụtara uto linear karịa mmụba nke a hụrụ na G. stearothermophilus mCFUs. Enweela mkparịta ụka ogologo oge 52 gbasara ọdịdị nke ọnụ ọgụgụ nke mkpụrụ ndụ: ebe ụfọdụ ọmụmụ na-akọ na ọnụ ọgụgụ nke microbes na-eto eto bụ nke nha nha ha (n'ịdị elu), ndị ọzọ na-egosi ọnụ ọgụgụ na-adịgide adịgide (linear ma ọ bụ bilinear). Dị ka Tzur et al.53 kọwara, ịmata ọdịiche dị n'etiti exponential na (bi) na-eto eto chọrọ kpọmkwem <6% na nha biomass, nke na-agaghị eru maka ọtụtụ usoro QPM, ọbụna gụnyere interferometry. Dị ka Tzur et al.53 kọwara, ịmata ọdịiche dị n'etiti exponential na (bi) na-eto eto chọrọ kpọmkwem <6% na nha biomass, nke na-agaghị eru maka ọtụtụ usoro QPM, ọbụna gụnyere interferometry. Как OBъяснил Цур и др.53, различение ? остижимо для большинства методов QPM, даже с использованием интерферометрии. Dị ka Zur et al.53 kọwara, ịmata ọdịiche dị n'etiti exponential na (bi) na-eto eto chọrọ <6% izi ezi na nha biomass, nke a na-apụghị iru maka ọtụtụ ụzọ QPM, ọbụna na-eji interferometry.Dị ka Zur et al kọwara. 53, ịmata ọdịiche dị n'etiti exponential na (bi) uto linear chọrọ ihe na-erughị 6% izi ezi na nha biomass, nke enweghị ike iru maka ọtụtụ ụzọ QPM, ọbụlagodi mgbe ejiri interferometry. CGM na-enweta izizi nke a site na izi ezi sub-pg na nha biomass36,48.
Oge nkwụsị nke nwere onyonyo 6 OT (mikpu mmanụ, 60x, ebumnobi NA 1.25) na (b) evolushọn micro-CFU biomass tụrụ na CGM. Lee ihe nkiri M7 maka ozi ndị ọzọ.
Uto ahịrị ahịrị zuru oke nke S. shibatae bụ ihe a na-atụghị anya ya na a kọbeghị ya. Otú ọ dị, a na-atụ anya ito eto, ma ọ dịkarịa ala n'ihi na ka oge na-aga, ọtụtụ nkewa nke 2, 4, 8, 16 ... ga-emerịrị. Anyị chere na uto ahịrị nwere ike ịbụ n'ihi mgbochi cell n'ihi mkpọkọta mkpụrụ ndụ buru ibu, dịka uto cell na-ebelata ma mesịa rute ọnọdụ ụra mgbe njupụta cell dị oke.
Anyị na-ekwubi site n'ịtụle isi ihe ise ndị a na-adọrọ mmasị n'otu n'otu: Mbelata nke ọkụ ọkụ, Mbelata ọkụ ọkụ, mmasị na ọla edo nanoparticles, mmasị na quantitative frasi microscopy, na a ga-ekwe omume okpomọkụ nso nke LA-HTM nwere ike iji.
E jiri ya tụnyere ikpo ọkụ na-eguzogide, ọkụ laser ejiri maka mmepe HTM na-enye ọtụtụ uru, nke anyị gosipụtara n'ọmụmụ ihe a. Karịsịa, na mgbasa ozi mmiri mmiri n'ọhịa nke microscope, a na-edobe olu kpo oku n'ime mpịakọta 3 ole na ole (10 μm). N'ụzọ dị otú a, ọ bụ nanị ụmụ nje ndị a na-ahụ anya na-arụ ọrụ, ebe nje bacteria ndị ọzọ na-ehi ụra ma nwee ike iji ya mee ihe n'ihu na-amụ ihe atụ - ọ dịghị mkpa ịgbanwe ihe nlele ahụ mgbe ọ bụla ọ dị mkpa ka a nyochaa okpomọkụ ọhụrụ. Tụkwasị na nke ahụ, ikpo ọkụ microscale na-enye ohere nyocha kpọmkwem nke nnukwu okpomọkụ: A na-enweta ihe oyiyi 4c site na ihe nkiri 3-hour (Movie M3), nke na-achọkarị nkwadebe na nyocha nke ọtụtụ ihe atụ - otu maka nke ọ bụla n'ime ihe ndị a na-amụ. y bụ okpomọkụ na-anọchite anya ọnụọgụ ụbọchị na nnwale ahụ. Mbelata ụda dị ọkụ na-edokwa ihe niile dị gburugburu anya nke microscope, ọkachasị oghere ebumnobi, na okpomọkụ ụlọ, nke bụ nnukwu nsogbu ndị obodo chere ihu ruo ugbu a. Enwere ike iji LA-HTM na oghere ọ bụla, gụnyere lenses imikpu mmanụ, ọ ga-anọkwa na ọnụ ụlọ okpomọkụ ọbụlagodi na oke okpomọkụ n'ọhịa nlele. Isi njedebe nke usoro ikpo ọkụ laser nke anyị na-akọ na ọmụmụ ihe a bụ na mkpụrụ ndụ ndị na-adịghị agbaso ma ọ bụ na-ese n'elu mmiri nwere ike ịdị anya site na nlele anya ma sie ike ịmụ ihe. Ihe na-arụ ọrụ nwere ike ịbụ iji lenses na-ebuwanye ibu dị ala iji nweta ịrị elu okpomọkụ ka ukwuu karịa narị micron ole na ole. A na-eso ịdọ aka ná ntị a na mbelata mkpebi oghere, ma ọ bụrụ na ihe mgbaru ọsọ bụ ịmụgharị mmegharị nke microorganisms, ọ dịghị achọ mkpebi dị elu.
Ogologo oge maka kpo oku (na jụrụ oyi) usoro \({{{{\rm{\tau}}}}}}}_{{{\mbox{D}}}}\) dabere na nha ya, dị ka iwu \ ({{{({\rm{\tau }}}}}}_{{\mbox{D}}}={L}^{2}/D\), ebe \ (L \) bụ njirimara nha nke isi iyi ọkụ ( dayameta nke eriri laser na ọmụmụ anyị bụ \ (L \ banyere 100 \) μm), \ (D \) bụ diffusivity nke gburugburu ebe obibi (nkezi na anyị). ikpe, iko na mmiri Diffusion ọnụego\(D\ banyere 2\fold {10}^{-7}\) m2/s). Mgbanwe ọnọdụ okpomọkụ, enwere ike ịtụ anya nguzobe ọnọdụ okpomọkụ a na-eme ngwa ngwa ọ bụghị nanị na-ebelata oge nnwale ahụ, kamakwa na-enye ohere oge kpọmkwem \(t=0\) maka ọmụmụ ihe ọ bụla siri ike nke mmetụta okpomọkụ.
Usoro anyị tụrụ aro na-adabara na mkpụrụ osisi ọ bụla na-enye ọkụ (dịka ọmụmaatụ, ihe nlere azụmaahịa nwere mkpuchi ITO). Otú ọ dị, ọla edo nanoparticles na-enwe ike inye nnukwu absorption na infrared na obere absorption na nso nso a na-ahụ anya, njirimara nke ikpeazụ bụ nke na-adọrọ mmasị maka nlele anya anya dị irè na ebe a na-ahụ anya, karịsịa mgbe ị na-eji fluorescence. Na mgbakwunye, ọla edo bụ ihe dakọtara, kemịkalụ inert, njupụta anya nwere ike gbanwee site na 530 nm ruo nso infrared, na nkwadebe nlele dị mfe na akụnụba29.
Transverse grating wavefront microscopy (CGM) na-enye ohere ọ bụghị naanị eserese okpomọkụ na microscale, kamakwa nlekota biomass, na-eme ka ọ baa uru karịsịa (ma ọ bụrụ na ọ dịghị mkpa) yana LA-HTM. N'ime afọ iri gara aga, e mepụtala usoro microscopy okpomọkụ ndị ọzọ, karịsịa n'ihe gbasara bioimaging, na ọtụtụ n'ime ha chọrọ iji nyocha fluorescent nke nwere mmetụta okpomọkụ54,55. Otú ọ dị, a katọrọ ụzọ ndị a na ụfọdụ akụkọ atụlewo mgbanwe okpomọkụ na-ezighị ezi n'ime sel, ikekwe n'ihi na fluorescence na-adabere n'ọtụtụ ihe na-abụghị okpomọkụ. Tụkwasị na nke ahụ, ọtụtụ nyocha nke fluorescent na-ejighị n'aka na okpomọkụ dị elu. Ya mere, QPM na karịsịa CGM na-anọchite anya usoro microscopy okpomọkụ dị mma maka ịmụ ndụ na okpomọkụ dị elu site na iji anya microscopy.
Nnyocha nke S. shibatae, nke na-ebi ndụ kacha mma na 80 ° C, na-egosi na LA-HTM nwere ike itinye n'ọrụ iji mụọ hyperthermophiles, ọ bụghị naanị thermophiles dị mfe. N'ụkpụrụ, enweghị oke na oke okpomọkụ nke enwere ike iru site na iji LA-HTM, na ọbụna okpomọkụ dị elu karịa 100 Celsius nwere ike iru na nrụgide ikuku na-enweghị isi, dị ka ndị otu 38 anyị gosipụtara na ngwa kemịkalụ hydrothermal na ikuku. nrụgide A. A na-eji laser eme ihe maka ikpo ọkụ nanoparticles ọla edo 40 n'otu ụzọ ahụ. Ya mere, LA-HTM nwere ike iji mee ihe iji hụ hyperthermophiles na-enwetụbeghị ụdị ya nwere ọkọlọtọ nyocha anya dị elu n'okpuru ọnọdụ ọkọlọtọ (ya bụ n'okpuru nrụgide gburugburu ebe obibi).
Emere nnwale niile site na iji microscope arụrụ n'ụlọ, gụnyere ọkụ ọkụ Köhler (ya na LED, M625L3, Thorlabs, 700 mW), ihe njide ihe atụ nwere mmegharị xy ntuziaka, ebumnuche (Olympus, 60x, 0.7 NA, ikuku, LUCPlanFLN60X ma ọ bụ 60x, 1.25 NA). , UPLFLN60XOI), CGM igwefoto (QLSI cross grating, 39 µm pitch, 0.87 mm si Andor Zyla igwefoto sensọ) iji nye ike na wavefront imaging, na sCMOS igwefoto (ORCA Flash 4.0 V3, 16-bit mode, si Hamamatsu) ka ịdekọ na data egosiri na onyonyo 5 (egwu mmiri nje). The dichroic beam splitter bụ 749 nm BrightLine onu (Semrock, FF749-SDi01). Nzacha n'ihu igwefoto bụ 694 mkpụmkpụ ngafe nzacha (FF02-694/SP-25, Semrock). Titanium sapphire laser (Laser Verdi G10, 532 nm, 10 W, pumped tsunami laser cavity, Spectra-Physics na Fig. 2-5, ọzọ dochie anya Millenia laser, Spectraphysics 10 W, pumped Mira laser cavity, Coherent, for Fig. 2 -5). 6 na 7) ka atọrọ n'okirikiri wavelength \({{({\rm{\lambda }}}}}}=800\) nm, nke dabara na plasmon resonance spectrum nke gold nanoparticles. Spatial light modulators (1920 × 1152 pikselụ) zụtara site na Meadowlark Optics gbakọọ hologram site na iji Gerchberg-Saxton algọridim dị ka akọwara na njikọ 39.
Cross grating wavefront microscopy (CGM) bụ usoro nyocha anya nke dabere na ijikọta grating diffraction akụkụ abụọ (nke a makwaara dị ka cross grating) n'ebe dị anya nke otu millimita site na ihe mmetụta igwefoto. Ihe atụ kachasị nke CGM nke anyị ji mee ihe n'ọmụmụ ihe a bụ nke a na-akpọ interferometer transverse shift interferometer anọ (QLSI), ebe cross-grating na-agụnye ihe siri ike / usoro checkerboard ụkpụrụ ewepụtara na patent nke Primot et al. na 200034. The vetikal na kehoraizin grating ahịrị na-emepụta okporo-dị ka onyinyo na ihe mmetụta, distortion nke nwere ike numerically hazie na ozugbo iji nweta ngwa anya wavefront distortion (ma ọ bụ Ẹkot na-adọ profaịlụ) nke merenụ ìhè. Mgbe ejiri ya na mikroskopu, igwefoto CGM nwere ike igosipụta ọdịiche ụzọ anya anya nke ihe onyonyo, nke a makwaara dị ka omimi anya (OT), yana nghọta na usoro nanometers36. N'ihe CGM ọ bụla, iji wepụ ntụpọ ọ bụla dị na ngwa anya ma ọ bụ beams, a ga-ewererịrị ma wepụ ihe oyiyi OT bụ isi na foto ọ bụla na-esote.
A na-eme microscopy okpomọkụ site na iji igwefoto CGM dịka akọwara na ntụnye aka. 32. Na nkenke, ikpo ọkụ mmiri mmiri na-agbanwe index refractive ya, na-emepụta mmetụta lens thermal nke na-agbagọ ọkụ ọkụ ahụ merenụ. A na-eji CGM tụọ mgbagha ihu ebili mmiri a wee hazie ya site na iji deconvolution algọridim iji nweta nkesa okpomoku akụkụ atọ na ọkara mmiri mmiri. Ọ bụrụ na a na-ekesa nanoparticles ọla edo n'otu n'otu n'ime ihe nlele ahụ, enwere ike ịme eserese okpomọkụ na mpaghara na-enweghị nje bacteria iji mepụta ihe oyiyi ka mma, nke bụ ihe anyị na-eme mgbe ụfọdụ. Enwetara onyonyo CGM ntụaka na-enweghị kpo oku (na laser gbanyụọ) wee weghara ya n'otu ebe dị na onyonyo a na laser.
A na-enweta nha igwe kpọrọ nkụ site na iji otu igwefoto CGM ejiri maka onyonyo okpomọkụ. E nwetara onyonyo ntụaka CGM site n'ịkwagharị ngwa ngwa na x na y n'oge mkpughe dị ka ụzọ isi na-anata ihe ọ bụla inhomogeneity na OT n'ihi ọnụnọ nke nje bacteria. Site na onyonyo OT nke nje bacteria, enwetara biomass ha site na iji mkpokọta onyonyo n'akụkụ ebe ahọpụtara site na iji akụkụ nkebi nke Matlab arụrụ n'ụlọ (lee nkeji nke “koodu ọnụọgụ”), na-agbaso usoro akọwara na ref. 48. Na nkenke, anyị na-eji relation \(m={\alpha}^{-1}\int {{\mbox{OT}}\ekpe(x,y\right){{\mbox{d}} } x{{\mbox{d}}}y\), ebe \({{\mbox{OT}}}\ekpe(x,y\right)\) bụ onyinyo omimi anya, \(m\) bụ arọ akọrọ na \({{{{\rm{\alpha }}}}}\) bụ ihe na-adịgide adịgide. Anyị họọrọ \({{{\rm{\alpha)))))))=0.18\) µm3/pg, nke bụ ihe na-agbanwe agbanwe maka mkpụrụ ndụ dị ndụ.
A na-etinye ihe mkpuchi mkpuchi 25 mm na dayameta na 150 µm gbara ọkpụrụkpụ nke ejiri ọla edo nanoparticles kpuchie n'ime ụlọ AttofluorTM (Thermofisher) nke nwere nanoparticles ọla edo chere ihu. Geobacillus stearothermophilus emebere ya otu abalị na ọkara LB (200 rpm, 60°C) tupu ụbọchị ọ bụla nke nnwale. A na-etinye dobe nke 5 µl nke nkwusioru nke G. stearothermophilus nwere njupụta anya (OD) nke 0.3 ruo 0.5 n'elu mpempe akwụkwọ mkpuchi nke nwere nanoparticles ọla edo. Mgbe ahụ, a na-atụba ihe mkpuchi mkpuchi gburugburu 18 mm n'obosara nke nwere oghere 5 mm na dayameta dị n'etiti na dobe, na 5 μl nke nkwusioru nje na otu njupụta anya na-etinye ugboro ugboro na etiti oghere ahụ. A kwadebere olulu mmiri ndị dị na mkpuchi dịka usoro akọwapụtara na ref. 45 (lee Ozi mgbakwunye maka ozi ndị ọzọ). Mgbe ahụ tinye 1 ml nke ọkara LB na mkpuchi mkpuchi iji gbochie oyi akwa mmiri mmiri ka ọ ghara ikpochapụ. A na-etinye ihe mkpuchi ikpeazụ n'elu mkpuchi mechiri emechi nke ụlọ Attofluor™ ka ọ ghara ikpochapụ nke ọkara n'oge ntinye. Maka nnwale germination, anyị na-eji spores, nke, mgbe nyocha nke mbụ, mgbe ụfọdụ na-ekpuchi mkpuchi elu. A na-eji usoro yiri nke ahụ nweta Sulfolobus shibatae. Ụbọchị atọ (200 rpm, 75 ° C) nke izizi nke Thiobacillus serrata ka emere na ọkara 182 (DSMZ).
A kwadebere ihe atụ nke nanoparticles ọla edo site na micellar block copolymer lithography. A kọwara usoro a n'ụzọ zuru ezu na isi. 60. Na nkenke, a na-emepụta micelles na-ekpuchi ion ọla edo site na ịgwakọta copolymer na HAuCl4 na toluene. Mgbe ahụ, a na-emikpu ihe mkpuchi ndị a kpochara n'ime ngwọta ma mesoo ya na irradiation UV n'ihu onye na-ebelata ego iji nweta mkpụrụ ọla edo. N'ikpeazụ, a na-akụ mkpụrụ ọla edo site na ịkpọtụrụ mkpuchi mkpuchi nke nwere ngwọta mmiri nke KAuCl4 na ethanolamine maka nkeji iri na isii, nke butere nhazi oge na nke dị oke mma nke nanoparticles ọla edo na-abụghị okirikiri na infrared dị nso.
Iji tọghata interferogram ka ọ bụrụ ihe onyonyo OT, anyị na-eji algọridim arụrụ n'ụlọ, dịka akọwara ya na njikọ ahụ. 33 ma dị ka ngwugwu Matlab na ebe nchekwa ọha na-esote: https://github.com/baffou/CGMprocess. Ngwungwu a nwere ike gbakọọ ike yana onyonyo OT dabere na interferograms edekọtara (gụnyere onyonyo ntụaka) yana anya n'usoro igwefoto.
Iji gbakọọ usoro nke etinyere na SLM iji nweta profaịlụ okpomọkụ enyere, anyị jiri algọridim emebere emebere na mbụ nke dị na ebe nchekwa ọha na-esote: https://github.com/baffou/SLM_temperatureShaping. Ntinye bụ mpaghara okpomọkụ achọrọ, nke enwere ike ịtọ ya na dijitalụ ma ọ bụ site na onyonyo bmp monochrome.
Iji kewaa mkpụrụ ndụ ma tụọ ịdị arọ ha kpọrọ nkụ, anyị jiri Matlab algọridim bipụtara na ebe nchekwa ọha na-esote: https://github.com/baffou/CGM_magicWandSegmentation. Na onyonyo ọ bụla, onye ọrụ ga-pịa nje bacteria ma ọ bụ mCFU nke mmasị, mezie mmetụta wand, wee kwado nhọrọ ahụ.
Maka ozi ndị ọzọ gbasara imepụta ọmụmụ ihe, lee akụkọ nyocha nke okike ejikọtara na edemede a.
Data na-akwado nsonazụ nke ọmụmụ a dị site n'aka ndị odee dị iche iche na arịrịọ ezi uche dị na ya.
A kọwapụtara koodu isi mmalite a na-eme n'ọmụmụ ihe a na ngalaba Ụzọ, enwere ike ibudata nsụgharị debug na https://github.com/baffou/ na ebe nchekwa ndị a: SLM_temperatureShaping, CGMprocess, na CGM_magicWandSegmentation.
Mehta, R., Singhal, P., Singh, H., Damle, D. & Sharma, AK Insight n'ime thermophiles na ngwa ha sara mbara. Mehta, R., Singhal, P., Singh, H., Damle, D. & Sharma, AK Insight n'ime thermophiles na ngwa ha sara mbara.Mehta, R., Singhal, P., Singh, H., Damle, D. na Sharma, AK nkọwa nke thermophiles na ngwa ha sara mbara. Mehta, R., Singhal, P., Singh, H., Damle, D. & Sharma, AK 深入了解嗜热菌及其广谱应用。 Mehta, R., Singhal, P., Singh, H., Damle, D. & Sharma, AK.Mehta R., Singhal P., Singh H., Damle D. na Sharma AK Nghọta miri emi nke thermophiles na ọtụtụ ngwa ngwa.3 Biotechnology 6, 81 (2016).
Oge nzipu: Sep-26-2022