I-Hielscher Ultrasound Technology

Ukulungiselela kwe-Ultrasonic kwama-C. Elegans Amasampula

C. elegans, isibungu se-nematode, siyisidalwa semodeli esisetshenziswa kabanzi ku-biology. Ukulungiselela isampula ngaphambi kokuhlaziywa kudinga i-lysis, amaprotheni kanye ne-lipid extraction kanye nokuhlukaniswa kwe-RNA, okungenziwa ngokuthembekile nge-sonication. Iziphazamisi zeseli ze-Ultrasonic zithembekile, zisezingeni eliphezulu futhi zisebenziseka kalula ukulungiselela ngokushesha amasampula ka-elegans.

Ukulungiselela kwe-Ultrasonic kwama-C. Elegans Amasampula

C. elegans yizibungu eziyindilinga, ezisetshenziswa kakhulu kumalabhorethri ocwaningo ukuphenya ngezakhi zofuzo, i-biology yokuthuthuka nezifo. Izinhlobo eziningi zofuzo ku-C. elegans zinokufana okusebenzayo kubantu. Ngaleyo ndlela, isibungu se-nematode siyimodeli ewusizo kakhulu yezifo zabantu. Ezinye izinzuzo zokusetshenziswa okubanzi kwe-C. elegans ukutshalwa kwayo okulula nokushibhile emapuletini aqukethe amabhaktheriya (isb., E. coli), ukusebenza kwawo obala, ukuphatha okulula, kanye nethuba lokubamba nokugcina izikelemu isikhathi eside.
Ukuhlaziywa kwamaprotheni kanye ne-lipid kuyizinqubo ezijwayelekile emalabhorethri kanye nokulungiswa kwesampula kwe-ultrasonic kuyindlela esungulwe yokwenza i-lyse C. elegans nematodes kuzo zonke izigaba zokuthuthuka (okungukuthi imibungu, izibungu L1-L4, abantu abadala). Njengoba uC. Elegans ubuye asetshenziswe njengohlelo lokuveza amaprotheni ukuveza ngokweqile amaprotheni ahlosiwe, indlela ethembekile, ekhiqizwayo yokukhiqiza i-lysis kanye namaprotheni, enikeza isivuno esikhulu samaprotheni, iyadingeka. Izinhlelo zokuphazanyiswa nokukhishwa kwamaseli e-Ultrasonic ziyatholakala njengama-homogenizers wohlobo lwe-probe futhi njengama-ultrasonicators wesampula amaningi. Ukuhlinzekela ukulungiselela okulula kwesampula kanye nalo lonke uhlobo lwezinombolo zosayizi wesampula, i-Hielscher Ultrasonics inesiphazamisi esihle se-ultrasonic cell senqubo yakho yelabhu.
C. elegans is a widely used model organism in biological research. Ultrasonic lysis is a sophisticated, reliable, reproducible and rapid technique to prepare high-quality protein extracts from C. elegans.

I-Ultrasonic C. elegans Lysis ye

  • Ukulungiselela homogenates worm
  • Izakhi zeprotheyini
  • Ukukhishwa kwe-lipid
  • Ukulinganiswa kwamaprotheni
  • Ukwehliswa komzimba
  • I-Western blotting
  • Ukukhishwa kwe-RNA
  • Ukuhlolwa kwe-Enzymatic
I-sonotrode MTP-24-8-96 inezinqubo eziyisishiyagalombili ze-ultrasonic zokunikezwa kwemithombo yamapuleti ama-microtiter.

I-sonotrode MTP-24-8-96 inezinqubo eziyisishiyagalombili ze-ultrasonic zokunikezwa kwemithombo yamapuleti ama-microtiter.

Isicelo solwazi




Qaphela kwethu Inqubomgomo yobumfihlo.


Izivumelwano ze-Ultrasonic zeC. Elegans Disruption and Lysis

I-Ultrasonic homogenization kanye ne-lysis ka-C. elegans kanye neprotheyini elandelayo kanye nokukhishwa kwe-lipid kungenziwa kusetshenziswa izinqubo ezahlukahlukene kusetshenziswa i-homogenization kanye ne-lysis buffers njll. Ngezansi, sikunikeza izivumelwano ezinokwethenjelwa nezisheshayo ze-ultrasonic lysis kanye nezindlela zokukhipha ukulungiswa kwamasampuli we-elegans aphezulu we-protein- noma i-lipid.

Izinzuzo ze-Ultrasonic C. elegans Lysis

  • ethembekile
  • i-reproducible
  • ilawulwa ngamazinga okushisa ngqo
  • inqubo ethembekile yokulawula
  • indlela emnene
  • kulula ukuyisebenzisa
  • ephephile

Isizinda se-Ultrasonic Protein kusuka ku-C. elegans Amasampula

I-Ultrasonic lysis kanye nokukhishwa kwamaprotheni ka-C. elegans worms kungenziwa kusetshenziswa izivumelwano ezahlukahlukene. Ngezansi sikwethulela izinqubo ezimbalwa ezinokwethenjelwa nezisheshayo ze-lysis zemiphumela yokukhipha amaprotheni.

Ukulungiswa Ngokushesha Kwe-Cytosolic Extract kusuka ku-C. elegans Izikelemu nge-Sonication

Ngephrothokholi elandelayo ungalungiselela uC. Elegans lysates ngemizuzu engaphansi kwengu-30.
Ukuqoqwa kwama C. elegans
Khetha izibungu ze-elegans ozifunayo zibe yi-1.5ml tube phosphate buffered saline (PBS) noma uzigeze kupuleti nge-1.5ml PBS. ICentrifuge ngo-1min ngo-2000rpm kuya ku-pellet. Gcina amasampuli ngaso sonke isikhathi eqhweni.
Ngemuva kwalokho, geza izimpethu kabili nge-PBS.
Ngemuva kwalokho, geza izimpethu kabili nge-ddH2O.
Phinda usebenzise izimpethu okungenani ku-500ul we-homogenization buffer (HB). Amasampuli ezikelemu manje aselungele i-ultrasonic lysis.
Ukuze uthole ikhwalithi ephezulu yokukhipha amaprotheni, ungahle uthande ukunciphisa ukungcoliswa kwamagciwane ngokugeza izikelemu ze-5min lilinye ku-PBS nakumanzi oyinyumba, angenawo amandla (ddH2O) noma wenze i-sucrose floatation. Gcina amasampuli ezikelemu ngokuqhubekayo eqhweni.

I-Ultrasonic C. elegans Lysis Protocol

  • Qiniseka ukuthi ulungiselela i-ultrasonicator ngaphambili ukuze i-homogenizer ye-ultrasonic ilungele ukusetshenziswa (ukukhishwa kwephenyo, uhlelo lwe-sonication pre-set).
  • Ultrasonicator UP200Ht (200 watts, 26kHz) with microtip S26d2 for the preparation of biological samplesUma i-ultrasonicator yakho imiswe kahle, beka i-ice-bath ngamashubhu wakho wesampula ngaphansi kwephenyo le-ultrasonic bese ufaka iphenyo le-ultrasonic kuthubhu engu-1.5ml.

  • Ku-C. elegans lysis ne-UP200St noma i-UP200Ht, i-ultrasonication kufanele yenziwe kusetshenziswa i-microtip (isb. 2mm probe S26d2; bheka isithombe kwesobunxele) ku-40% amplitude ye-1sec enama-30sec ikhefu phakathi. Imijikelezo engu-5 ye-sonication kumzuzwana ngamunye ongu-1 onamakhefu angama-30sec ilungele uC elegans lysis. Uma wenza i-lysis okokuqala ngqa, ungabheka ukuqhubekela phambili kwe-lysis kuma-aliquots amancane esampuli ngemuva kokudonsa ngakunye usebenzisa imicroscope.
  • ILysis igcwaliswa ngempumelelo lapho izimpethu ziphazamiseka. I-over-sonication iholela ekwehlukaneni kwe-nuclei futhi ibonakale lapho isampuli ithola i-viscous noma amagwebu. Ukuvikela ukonakala kwesampula, sebenzisa ama-pulses amaningi uma kudingeka. Musa ukwandisa isikhathi somjikelezo ngamunye we-ultrasonic pulse ukuthola ama-protein aphezulu.
  • Sula i-cell lysate ngokufaka phakathi izikelemu ezine-lysed nge-14,000rpm nge-10min ku-4ºC.
  • Ngemuva kwalokho, thumela isidalwa esingaphezu kwesishubhu kushubhu elisha bese ulungiselela ukuncishiswa komzimba noma okunye ukuhlolwa.

Inothi le-homogenization buffer: Lungiselela i-homogenization buffer ye-ultrasonic lysis protocol engenhla ngokulandelayo:

  • I-15 mM Hepes pH 7.6 – 15 ml ka-0.5 M
  • 10 mM KCl – 2.5 ml ka-2 M
  • 1.5 mM MgCl2 – 0.75 ml ka-1 M
  • 0.1 mM EDTA – I-100 ul engu-0.5 M
  • 0.5 mM EGTA – 2.5 ml we-0.1 M
  • 44 mM iSucrose – 14.7 ml ka-50%
  • Faka ngaphambi kokusebenzisa: 1mM DTT – I-1000x ye-1M
  • kanye ne-protease inhibitor

I-Ultrasonic Lysis ka-C. elegans ye-Quantitative Affinity Yokuhlanzwa Izilingo

C. imibungu ye-elegans (∼2 million replicate) isanda kuvunwa ku-triplicate ye-biological ngokufaka i-hermaphrodites encane ye-gravid futhi yenziwe i-ice (umjikelezo: 0.5 s, amplitude: 40-45%, 5 strokes / session, 5 sessions, interval between sessions : Amashumi amathathu; Iprosesa ye-UP200S ye-ultrasonic nge-micro-tip S26d2 (Hielscher Ultrasonics GmbH)) ku-lysis buffer (ivolumu ephelele: ∼600 μl; 50 mm Tris-HCl, pH 7.4, 100 mm KCl, 1 mm MgCl2, 1 mm EGTA, 1 mm DTT, 10% glycerol , ingxube ye-protease inhibitor, i-0.1% ye-Nonidet P-40 Substitute). Ngemuva kwe-sonication, i-Nonidet P-40 Substitute yengezwe yafika ku-1% kanti ama-lysates afakwa ekhanda phezu komsila ngokujikeleza ku-4 ° C ngemizuzu engama-30, kwalandelwa yi-centrifugation ngo-20,000 × g ngemizuzu engama-20 ngo-4 ° C. I-lysate esuliwe yabe isifiswa ngaphandle kokuphazamisa ungqimba olungaphezulu lwe-lipid futhi yahlukaniswa ngesigamu yaba ubuhlalu be-anti-GFP agarose noma ubuhlalu bokulawula obuvinjiwe (40-50 μl). Ngemuva kokujikisa ikhanda ngomsila ku-4 ° C ngamaminithi angama-60-90, ubuhlalu bugezwe kanye nge-lysis buffer equkethe i-0.1% Nonidet P-40 Substitute, kwalandelwa izikhathi ezimbili zokuwasha kunoma iyiphi i-buffer I (25 mm Tris-HCl, pH 7.4, 300 mm NaCl, 1 mm MgCl2) noma i-buffer II (1 mm Tris-HCl, pH 7.4, 150 mm NaCl, 1 mm MgCl2) noma zombili. Okwe-GFP: MBK-2 donsa-phansi, izivivinyo ezimbili ezihlukene zenziwa kusetshenziswa izimo ezahlukahlukene zokuwasha. Amaprotheni ahlonishwa ngokuzamazama kwe-orbital ku-50 μl ka-6 m urea / 2 M thiourea ekamelweni lokushisa. Okocwaningo lokudonswa kwe-MBK-1 :: GFP, amaprotheni ahlonishwa kabili ngokuzamazama ku-50μl ka-8 m we-guanidinium chloride ku-90 ° C, kwalandelwa imvula ye-ethanol. Amasampula amaprotheni ahlanganisiwe abe esegaywa esisombululo.
(Bheka uChen et al., 2016)

VialTweeter at the ultrasonic processor UP200ST

I-VialTweeter iyunithi yokulungiselela amasampula amaningi wokwehlukaniswa kanyekanye kwamashubhu wokuhlola ayi-10.

I-Ultrasonic Worm Homogenization neLysis

Ngenqubo ye-C.elegans lysis nenqubo yokukhipha amaprotheni, kwaqoqwa ama-nemotode angama-30,000 esiteji esifanelekile futhi agezwa ku-S-basal ebanda kakhulu eqhweni, kugxilwe yi-centrifugation ngo-1500 rpm ngemizuzu emi-2., Wageza amahlandla ayisithupha nge-ice-cold S- i-basal ukususa amagciwane asalayo, bese igcinwa eqhweni ize ilungele ukusetshenziswa. Ngokukhishwa kwamaprotheni, izibungu ze-gravid-omdala zavunyelwa ukuthi zenze i-compact pellet ngemuva kokuwashwa kokugcina kwe-S-basal. Ama-pellets ama-worm abuye avuselelwa ku-1 ml we-Ice-cold Extraction Buffer [20 mM potassium phosphate, pH 7.4, 2mM EDTA, 1% Triton-X-100, protease inhibitors (Sigma P2714)] acutshungulwa ngokushesha.
Izimpethu ezingama-gravid-omdala ezingama-30,000 (ezihambelana nesisindo esingu- ∼100 mg esimanzi), zenziwe zanikwa iqhwa kusetshenziswa i-probe-type ultrasonicator (isb. 1 ml we-ice-cold extraction buffer. (Bheka uBaskharan et al. 2012)

Isizinda se-Ultrasonic Lipid esivela ku-C. elegans

Ku-lipidomics, igatsha le-metabolomics, ukuhlanganiswa kwe-lipid kwezinhlelo zebhayoloji kuyabonakala futhi kuyahlaziywa. C. elegans asetshenziswa kakhulu kuma-lipidomics ukuphenya ukusebenzisana kwama-lipids we-metabolic nemiphumela yawo empilweni nasempilweni.
I-Ulysis ye-Ultrasonic kanye nesizinda kusetshenziselwa ukukhipha ama-lipids afana ne-sphingolipids avela ku-C. Elegans imibungu, izibungu, nezimpethu ezindala. I-Ultrasonication isetshenziselwa ukulungiselela ama-homogenates wesibungu futhi ngemuva kwalokho kukhishwe ama-lipids kusuka kusampula.
I-Protocol ye-Ultrasonic Lipid Extraction evela ku-C. elegans
Faka i-elegans pellet eqhweni bese uphinde uyisebenzise nge-0.5 ml ultrawater. Hlanganisa amasampula e-elegans ku-1.5mL amashubhu wokuhlola, ngenkathi ugcina amasampula ngokuqhubekayo eqhweni.
I-Sonication ingenziwa kusetshenziswa i-probe-ultrasonicstor efana ne- UP200Ht, iyunithi yesampula yokulungiselela isampula I-VialTweeter (sonication kanyekanye yamasampuli ayi-10) noma i- I-UIP400MTP (i-sonication yamapuleti amaningi anama-plate angama-96). Nge-lysis ye-ultrasonic ene-UP200Ht sebenzisa i-micro-tip S26d2. Hlela kuqala imodi yomjikelezo we-ultrasonic kwimenyu yedijithali. Setha i-amplitude ibe yi-10% nemodi yomjikelezo we-sonication yamapulse ama-2 sec wemijikelezo engama-20 ngekhefu lama-30 sec. phakathi kokuqhuma ngakunye kwe-ultrasonic pulsation.
Dlulisa ama-supernatants kumashubhu engilazi nge-screw cap. Yenza isizinda se-Folch ngokungeza i-1 ml ultrawater kuthubhu ngayinye yengilazi, kulandelwe ngokungeza u-6 ml wenhlanganisela ye-chloroform / methanol (ratio = 2: 1) kuthubhu ngayinye yengilazi.
I-Vortex ithubhu ngayinye yengilazi amasekhondi angama-30 izikhathi ezi-4. ICentrifuge amashubhu ku-1,258 xg ngemizuzu engu-15 (i-Eppendorf, 5810 R) ukuqhubekisela phambili ukuhlukaniswa kwesigaba. Dlulisa ingxenyenamba engezansi ye-hydrophobic kushubhu yengilazi ehlanzekile ngepayipi lengilazi iPasteur. Yomisa ingxenye engezansi ye-hydrophobic ngaphansi kokugeleza kwe-nitrogen ku-evaporator ye-nitrogen. Gcina i-pellet eyomile efrijini -80 ° C kuze kube yilapho uyisebenzisa.

Ukulungiselela i-Ultrasonic kwe-Worm Lysates

I-Wys lysate: Kwavunwa izikelemu zesigaba se-L4 zagezwa kathathu nge-M9 buffer (42.26 mM Na2I-HPO4, 22.04 mM KH2PO4, 85.56 mM NaCl, kanye no-0.87 mM MgSO4) ukuze ususe wonke amabhaktheriya. Ngemuva kokukhipha okuningi ngangokunokwenzeka kwe-M9 buffer, izimpethu zaphinde zathunyelwa ku-lysis buffer: 50 mM HEPES, 50 mM KCl, 1mM EDTA, 1mM EGTA, 5 mM phosphate β-glycerol, 0.1% (v / v) Triton X-100, 50mM sodium fluoride, 1 mM sodium orthovanadate, 5 mM sodium pyrophosphate, 0.2 mM phenylmethanesulfonylfluoride ne-protease inhibitor. Izikelemu zafriziwa ku-nitrogen ewuketshezi zancibilika ku-37 ° C amahlandla amathathu, khona-ke izibungu zanikelwa eqhweni elomile ngeyunithi ye-VialTweeter ye-ultrasonic yokulungiselela ngasikhathi sinye amashubhu wesampula ayi-10. I-Sonication yenziwa ngo-50% amplitude ngemijikelezo eyi-10 yamasekhondi ama-2. ngokuma okwesikhashana okungu-30 phakathi kokuqhuma kwe-sonication. Ngemuva kwalokho, amasampuli afakwa phakathi nge-12000 rpm ku-4 ° C ngemizuzu eyi-15. I-supernatant yaqoqwa yagcinwa ku -70 ° C. I-aliquot yasetshenziselwa ukulinganisa amaprotheni ngesilingo sikaBradford.
Ukunqunywa kwe-glutathione ephelele, i-GSH, ne-GSSG: Ngokulinganiswa kwe-glutathione, ama-lysates nokuzimisela kwenziwa ngalo lolo suku. Izibungu ezondliwe ngeglucose nokulawula i-L4 zavunwa zagezwa nge-M9 buffer kathathu. Ngemuva kokukhipha i-M9 buffer ngangokunokwenzeka, izikelemu zaphinde zathunyelwa ku-ice-cold metaphosphoric acid (5% w / v), izikelemu zadluliselwa eqhweni nge-VialTweeter ye-ultrasonic ku-50% amplitude kumijikelezo eyishumi ye-sonication yamasekhondi amabili. namasekhondi angama-30 phumula phakathi komjikelezo ngamunye. Ngemuva kwalokho, kufakwe phakathi ku-12000 rpm ngo-4 ̊C ngamaminithi ayi-15.
(cf Alcántar-Fernández et al., 2018)

C. elegans Sample Prep before Immunoprecipitation and Western Blotting

Ngamafuphi, ngokukhishwa kombungu, uC. elegans L1 izibungu zakhuliswa ngamasiko amakhulu we-S-medium ukuba abe mdala. Imibungu yaqoqwa kusetshenziswa indlela ejwayelekile ye-bleach futhi yamiswa ku-lysis buffer (50 mM Tris, pH 7.5, 100 mM KCl, 1 mM EDTA, 1 mM MgCl2, 8.7% glycerol, 0.05% NP-40, 1􏰅 Protease Inhibitor Cocktail, kanye 1􏰅 Phosphatase Inhibitor Cocktail I no-II), iqandiswe masishane ku-nitrogen ewuketshezi, futhi ifakwe lysed yi-ultrasonic cell ukuphazanyiswa kusetshenziswa i-ultrasonicator ene-microtip efana ne UP200Ht nge-S26d2 ngama-pulses ayi-10 ngaphezulu kwe-10 s kuma-30% amplitude. Uma inani elikhulu lamasampuli kufanele lilungiselelwe i-ultrasonic I-VialTweeter noma i-MultiSample-Ultrasonicator UIP400MTP yamapuleti kahle iyanconywa. Ngemuva kwe-sonication, okukhishwe kususwe ngaphambili nge-centrifugation ku-30,000g ngemizuzu engama-20 ku-4 ° C. Ukukhishwa okusulwe ngaphambili (i-300µg yamaprotheni esewonke) kufakwe ne-40µ􏰂g ye-anti-CDC-25.1 anti-purified antibody (lolu cwaningo) enxhunyaniswe ne-protein A-agarose, noma njengokulawula, inani elifanayo le-rabbit immunoglobulin (Ig) G enqamulelwe ukuxhunyaniswa ne-protein A-agarose isetshenziswe kumthamo ophelele we-200µl we-lysis buffer equkethe i-1% NP-40, ukufakwa kwayo okunciphise ukubopha okungacaciswanga kwamaprotheni ku-matrix. Amasampuli azungeziswa i-1 h ku-4 ° C, ubuhlalu bugezwe kathathu nge-lysis buffer, futhi bahlanganiswa ne-30µl ye-glycine / HCl ne-200 mM NaCl, pH 2.2. Ngemuva kokuncishiswa komzimba, ama-eluates ahlanjululwa ngo-30µ􏰂l wesampula yesampula ye-SDS, afudunyezwa aya ku-95 ° C ngemizuzu emi-4, futhi imvamisa u-3% wenani lokufaka ne-30% yama-elues asetshenzisiwe ku-SDS-PAGE elandelwa ukuvimba kwe-Western nge-anti -CDC-25.1 (1: 400), anti-LIN-23 (1: 750), anti-ubiquitin (1: 1000), anti-GSK3􏰁 (1: 500), noma anti-􏰁β-actin (1: 2000) amasosha omzimba. Lapho okukhishwe kungabanjiswanga ukugonywa komzimba, inani elifanayo lamaprotheni aphelele atholakala kulezo zinto ezikhishwe laphinde lafakwa kusampula le-SDS, lafudunyezwa laya ku-95 ° C, lase lisetshenziswa ngqo ku-SDS-PAGE futhi lahlaziywa yi-Western blotting. (Bheka uSegref et al. 2020)

I-probe-type insonifier UP200St ye-lysis

Isiphazamisi seseli se-Ultrasonic UP200St nge-micro-tip S26d2 yokukhishwa kwe-lysis kanye namaprotheni

I-Ultrasonic Lysis ngaphansi kwe-Prescise Temperature Control

The VialTweeter is a MultiSample Ultraonicator that allows for reliable sample preparation under precisely controlled temperature conditions.Ukulawulwa okuqondile nokunokwethenjelwa kokushisa kubalulekile lapho usingatha amasampula wezinto eziphilayo. Amazinga okushisa aphezulu aqala ukonakaliswa kwamaprotheni okubangelwa ukushisa ngamasampula.
Njengawo wonke amasu wokulungiselela isampula, i-sonication idala ukushisa. Kodwa-ke, izinga lokushisa lamasampuli lingalawulwa kahle lapho usebenzisa i-VialTweeter. Sikunikeza izinketho ezahlukahlukene zokuqapha nokulawula ukushisa kwamasampuli akho ngenkathi uzilungiselela nge-VialTweeter neVialPress ukuze zihlaziywe.

  1. Ukuqapha izinga lokushisa kwesampula: Iprosesa ye-ultrasonic i-UP200St, eshayela i-VialTweeter, ifakwe isoftware ehlakaniphile nenzwa yokushisa engaxhuma. Xhuma inzwa yokushisa ku-UP200St bese ufaka ichopho lenzwa yokushisa kwelinye lamashubhu esampula. Nge-digital-touch touch-display, ungasetha kumenyu ye-UP200St ibanga elithile lokushisa le-sonication yakho yesampula. I-ultrasonicator izoma ngokuzenzakalela lapho kufinyelela izinga lokushisa okuphezulu futhi imise isikhashana kuze kube yilapho izinga lokushisa lesampula lehla kunani eliphansi lokushisa okusethiwe ∆. Ngemuva kwalokho i-sonication iqala ngokuzenzekelayo futhi. Lesi sici esihlakaniphile sivimbela ukonakala okubangelwa ukushisa.
  2. Ibhlokhi ye-VialTweeter ingapholiswa ngaphambilini. Faka i-VialTweeter block (kuphela i-sonotrode ngaphandle kwe-transducer!) Efrijini noma efrijini ukupholisa ngaphambi kwebhulokhi ye-titanium kusiza ukuhlehlisa ukukhuphuka kwamazinga okushisa kusampula. Uma kunokwenzeka, isampuli uqobo lwayo ingapholiswa ngaphambilini nayo.
  3. Sebenzisa iqhwa elomile ukupholisa ngesikhathi se-sonication. Sebenzisa ithileyi elingajulile eligcwele iqhwa elomile bese ubeka i-VialTweeter eqhweni elomile ukuze ukushisa kuphele ngokushesha.

Thola i-Optimal Ultrasonic Cell Disruptor yohlelo lwakho lwe-Lysis

I-Hielscher Ultrasonics ingumkhiqizi onesipiliyoni sesikhathi eside wokuphazamisa ukusebenza kwamaseli aphezulu we-ultrasonic kanye nama-homogenizers ama-laboratories, amasistimu ebhentshi phezulu kanye nezilinganiso zezimboni. Usayizi wesiko lakho leseli lebhaktheriya, umgomo wakho wocwaningo noma wokukhiqiza kanye nevolumu yeseli elizocubungulwa ngehora noma ngosuku yizinto ezibalulekile zokuthola ukuphazamiseka kweseli kwe-ultrasonic okulungile kohlelo lwakho lokusebenza.
I-Hielscher Ultrasonics inikeza izixazululo ezahlukahlukene zokunikezwa ngasikhathi sinye kwamasampuli amaningi (izitsha ezifika kwezingu-10) kanye namasampuli amaningi (okungukuthi, amapuleti ama-microtiter / amapuleti angama-96-kahle), i-classic probe-type lab ultrasonicator enamazinga ahlukene wamandla kusuka ku-50 kuye ku Ama-watts angama-400 kuma-processor we-ultrasonic asebenza ngokugcwele anama-16,000watts ngeyunithi ngayinye yokuphazamiseka kwamaseli ezentengiselwano kanye nokukhishwa kwamaprotheni ekukhiqizeni okukhulu. Onke ama-ultrasonicators ama-Hielscher akhelwe ukusebenza kwe-24/7/365 ngaphansi komthwalo ogcwele. Ukuqina nokuthembeka kuyizici ezisemqoka zamadivayisi wethu we-ultrasonic.
Onke ama-homogenizers edijithali e-ultrasonic afakwe isoftware ehlakaniphile, isibonisi sokuthinta esinemibala kanye neprothokholi yedatha ezenzakalelayo, eyenza idivaysi ye-ultrasonic ibe yithuluzi lomsebenzi elilula kumalab nezindawo zokukhiqiza.
Sazise, luhlobo luni lwamaseli, mthamo muni, unamaza amangaki nokuthi unaliphi ilitshe ukucubungula amasampula akho ezinto eziphilayo. Sizokuncoma ukuthi uphazamise amaseli we-ultrasonic ofanele kakhulu izidingo zakho zenqubo.

Ithebula elingezansi likunikeza inkomba yamandla wokucutshungulwa alinganiselwe amasistimu wethu we-ultrasonic kusuka kuma-homogenizers aphethwe ngesandla nama-MultiSample Ultrasonicators kuya kuma-processor we-industrial ultrasonic wezinhlelo zokuthengisa:

Iqoqwana Ivolumu flow Rate Amadivayisi ezinconyiwe
Amapuleti angama-96-kahle / ama-microtiter na I-UIP400MTP
Izitsha eziyi-10 kuya ku-0.5 kuye ku-1.5mL na I-VialTweeter ku-UP200St
0.01 kuya ku-250mL 5 kuye ku-100mL / min UP50H
0.01 kuya ku-500mL 10 kuya ku-200mL / min UP100H
10 kuya ku-2000mL 20 kuya ku-400mL / min UP200Ht, UP400St
0.1 kuya ku-20L 0.2 kuya ku-4L / min I-UIP2000hdT
10 kuya ku-100L 2 kuya ku-10L / iminithi I-UIP4000hdT
na 10 kuya ku-100L / iminithi UIP16000
na sikhulu yinhlanganisela UIP16000

Xhumana nathi! / Cela Us!

Cela ulwazi oluthe xaxa

Uyacelwa ukuthi usebenzise ifomu elingezansi ukucela ulwazi olungeziwe mayelana nezinhlelo zokusebenza ze-ultrasonic, izinhlelo zokusebenza nentengo. Sizokujabulela ukuxoxa ngenqubo yakho nawe futhi sikunikeze uhlelo lwe-ultrasonic oluhlangabezana nezidingo zakho!









Sicela uqaphele wethu Inqubomgomo yobumfihlo.


Ultrasonic high-shear homogenizers are used in lab, bench-top, pilot and industrial processing.

I-Hielscher Ultrasonics ikhiqiza ama-homogenizers asebenza ngokusebenza okuphezulu wokuxuba izinhlelo, ukusabalalisa, ukumisa nokukhipha ilebhu, umshayeli kanye nesikali sezimboni.

Izincwadi / Izikhombo



Amaqiniso Okufanele Ukwazi

Ama-elegans e-Caenorhabditis

C. elegans yi-nematode ebonakalayo engafihli lutho (isibungu esiyindilinga) cishe eyi-1 mm ubude, edla amabhaktheriya (isb. E. coli) futhi inomjikelezo wempilo omfushane. Ku-20 ° C, uhlobo lwelabhorethri ka-C. elegans (N2) lunempilo yesilinganiso esimaphakathi kwamasonto ama-2-3 nesikhathi sesizukulwane sezinsuku ezi-3 kuya kwezi-4. Lapho ama-elegans ekhula ngobuningi, angenziwa kalula ngaphansi kwezimo zelebhu ezilawulwa ngokunembile, angahlolwa kalula ngomthetho osebenzayo wezidakamizwa zanoveli kanye nemiphumela yazo kanye nokusebenzisana ngaphakathi kwezinqubo eziyinkimbinkimbi zamangqamuzana ezifweni zomuntu. I-genome emfushane, umjikelezo wempilo emfushane nokusingathwa okulula kuzilungiselelo zelabhorethri kwenza uC. Elegans abe yimodeli enhle kakhulu yocwaningo efana ne-genomics, i-proteomics, i-biology yokuthuthuka, ucwaningo lwezifo, ukuthuthukiswa kwezidakamizwa njll.
I-Caenorhabditis elegans izikelemu kungaba ngamadoda noma i-hermaphrodite. Ama-Hermaphrodites anezitho zombili zokuzala zabesilisa nabesifazane. Kodwa-ke, izibungu zesifazane azikho. I-Hermaphrodites ingazimela noma ingazala nezibungu zesilisa. C. elegans angakhiqiza amaqanda angaphezu kwayi-1 000 nsuku zonke.
Njengoba uC. Elegans engesinye sezinto ezilula kakhulu ezinesistimu yezinzwa, isibungu se-nematode sisetshenziswa kusukela ngo-1963 njengesibonelo somzimba wocwaningo. Ama-neurons awashisi amandla esenzo, futhi awavezi noma yiziphi iziteshi ze-sodium ezinamandla. Ku-hermaphrodite, lolu hlelo luqukethe i-302 neuron iphethini yayo ebhalwe ngokuningiliziwe, kulokho okwaziwa njenge-connectome.
Izinhlobo eziningi zofuzo ku-C. elegans genome zinokufana okusebenzayo kubantu okwenza kube yimodeli ewusizo kakhulu ezifweni zabantu futhi ngokwesibonelo kusetshenziselwa ukutadisha i-biology yentuthuko, ukuguga nezinto ezithinta impilo ende. Ngaphezu kwalokho, uC. Elegans mutants uhlinzeka ngezibonelo zezifo eziningi zabantu kubandakanya ukuphazamiseka kwemizwa (isib. I-Alzheimer's), isifo senhliziyo esizelwe kanye nesifo sezinso.
Lezi zinto zenze uC elegans abe yimodeli ebaluleke kakhulu emikhakheni eminingi yocwaningo. Ngenxa yalokho, uC. Elegans waba yisidalwa sokuqala esinamaseli amaningi ukuthi lonke uchungechunge lwaso lulandelwe. I-genome iqukethe izinhlobo zofuzo ezilinganisa amaphesenti angama-20,470. Cishe ama-35% wezinhlobo zofuzo zika-elegans zinama-homologs angabantu. Ngokumangazayo, izakhi zofuzo zabantu zikhonjiswe kaninginingi ukuthi zithathe isikhundla sama-elegans homologs azo lapho zifakwa ku-C. elegans. Ngakolunye uhlangothi, izakhi zofuzo eziningi ze- C. elegans zingasebenza ngokufana nezakhi zofuzo ezincelisayo.

Isikhathi sempilo kaC. Elegans siseduze. Amaviki ama-3 futhi aqukethe izigaba zokuphila eziyisithupha: i-embryogenesis (isigaba samaqanda), izigaba ezine zesibungu (L1 kuya ku-L4), nesigaba sabadala. Ama-nematode aqanduselwa emaqandeni njengoba izibungu ze-L1 zinamaseli angama-560. Ukukhula ngesikhathi ngasinye sesigaba sesibungu kwenzeka ngokuhlukaniswa kwamaseli nangeseli hypertrophy. I-cuticular molting ibeka uphawu esigabeni ngasinye sesibungu. Uma izimo ezinzima zemvelo zikhomba kwisibungu esikhulayo ukuthi izimo akunakwenzeka ukuthi zisekele ukuzala kwabantu abadala, uC. Elegans angakushintsha ukukhula kwawo bese enza esinye isigaba sesibungu se-L3, lapho izibungu zingena esigabeni esinamandla. Kulesi simo, izilwane zibekezelela ngokweqile ukucindezeleka futhi zihlala isikhathi eside futhi zingaphila izinyanga ezintathu kuya kweziyisishiyagalolunye. Izibungu zeDauer ziyazihlukanisa nobunzima ngokufaka uphawu emigodini yazo ye-buccal ne-anal, zinciphisa amathumbu azo, futhi zivule uhlelo lwezofuzo oluxhomekeke ku-daf-16 / FOXO, phakathi kwezinye izinto, oluholela ekuvezweni kwe-cuticle eqondene ne-dauer. (bheka uHenderson et al., 2006)

C. elegans UDauer Larvae

Izibungu zeDauer yigama lezibungu ze-nematode, ezingene kwesinye isigaba sokuthuthuka. Igama elithi "izibungu zeDauer" lisetshenziselwa ikakhulukazi izibungu zomndeni wama-rhabditids kufaka phakathi ama-elegans eCaenorhabditis. Igama elithi "Dauer" livela eJalimane futhi lisho "ubude besikhathi” ngokwencazelo “isikhathi esithile ”. Izibungu zeDauer zingena ohlotsheni lwe-stasis futhi zingasinda ezimeni ezinzima. Uma futhi lapho isibungu singena esigabeni sokuqina sincike ezimweni zemvelo. Izibungu zeDauer zifundwa kabanzi kwi-biology ngoba izimbungulu zikhombisa amandla angajwayelekile okusinda ezindaweni ezinzima futhi ziphile isikhathi eside. Isibonelo, izibungu ze-C. elegans dauer zingaphila kuze kufike ezinyangeni ezine, isikhathi eside kakhulu kunesilinganiso seminyaka yokuphila kwazo cishe amasonto amathathu ngesikhathi sokukhula okuvamile kokuzala.