Kugwiritsa ntchito Artificial Cerebrospinal Fluid (ACSF) mu Patch Clamp Technology

Mawu osakira:Madzi Opanga Opangidwa ndi Cerebrospinal Fluid, ACSF, Artificial CSF, Synthetic cerebrospinal fluid, Synthetic CSF, CSF, CSF yoyeserera, Patch Clamp Technology

IPHASE Products

Mawu Oyamba

Kwa ofufuza a neurophysiology, Artificial Cerebrospinal Fluid (ACSF) ndi imodzi mwazinthu zofunika kwambiri komanso zomwe zimagwiritsidwa ntchito pafupipafupi pakuyesa kwa zigamba. M'malo mokhala yankho losasunthika, Artificial CSF imatanthawuza madera ovuta kwambiri omwe amatsimikizira kukhazikika, kuberekana, komanso kufunikira kwa thupi la kujambula kwa electrophysiological.

Cholinga cha kuyesa kwa patch clamp ndikutulutsanso, momwe kungathekere, momwe mu vivo magwiridwe antchito a ma neuron pansi pamikhalidwe ya ex vivo. Pakuyesa kumeneku, Artificial CSF imagwira ntchito ngati njira yothandizira. Kapangidwe kake ka ma ionic, kusungitsa mphamvu, osmolarity, ndi kufanana kwa mpweya zimakhudza mwachindunji kuthekera kwa neural resting membrane, kuwombera komwe kungachitike, kufalikira kwa ma synaptic, ndi machitidwe olowera njira.

The Physiological Basis of Native Cerebrospinal Fluid

Mfundo zonse zamapangidwe ndi ntchito za Artificial CSF zimamangidwa pakumvetsetsa kolondola kwa magwiridwe antchito amtundu wa cerebrospinal fluid.

Native cerebrospinal fluid ndi madzimadzi omveka bwino omwe amazungulira chigawo chapakati cha mitsempha, kupereka microenvironment yokhazikika yofunikira pazochitika za neuronal. Komabe, chifukwa cha kusiyanasiyana kwa kapangidwe kake komanso kupezeka kochepa, CSF yachibadwidwe siyingagwiritsidwe ntchito modalirika pazoyeserera zoyendetsedwa. Kuti athane ndi izi, Synthetic cerebrospinal fluid imapangidwa kuti ibwerezenso zofunikira zakuthupi za CSF, zomwe zimapangitsa ofufuza kukhalabe osasunthika, ogwirizana ndi maphunziro a in vitro ndi ex vivo.

Mtengo wa ACSF monga Essential Reagent mu Patch Clamp Experiments

M'mayesero a patch clamp, Artificial Cerebrospinal Fluid ndiye yankho lalikulu la extracellular. Ma neurons ndi ma cell a glial mkatikati mwa mitsempha yapakati amamizidwa mosalekeza mu CSF yakubadwa mu vivo, ndipo kuthekera kwawo kopumula kwa membrane, kuthekera kochitapo kanthu, ion channel kinetics, ndi kutulutsidwa kwa ma synaptic vesicle kumadalira kwambiri kukhazikika kwa ion, pH, osmolarity, ndi mphamvu.

Cholinga cha kujambula kwa patch clamp ndikutulutsanso mu vivo physiological state mokhulupirika momwe mungathere pansi pazikhalidwe za ex vivo. Monga kayeseleledwe kochita ka CSF, Artificial CSF imapereka malo okhazikika a minofu ndi ma cell, ndipo mumayendedwe ambiri a electrophysiology sizingasinthidwe ndi sing'anga wamba kapena saline wosavuta.

Mapulogalamu mu Patch Clamp Technology

Artificial CSF imagwiritsidwa ntchito panthawi yonse yoyeserera yachigamba, kuyambira pakukonzekera minofu mpaka kujambula ndikusintha kwamankhwala.

1. 1) Mayesero a Chigawo Chachikulu Chaubongo Patch Clamp

Mu ubongo dissection ndi slicing, minofu ayenera mofulumira kusamutsidwa mu ayezi-ozizira, mpweya ACSF (95% O₂ / 5% CO₂). Kutentha kwapang'onopang'ono kumathandizira kuchepetsa kagayidwe ka neuronal, pomwe kusanja bwino kwa gasi kumasunga kukhazikika kwa pH ndikuchepetsa kuwonongeka kwa hypoxic.

Pambuyo podula, zigawo za ubongo zimayikidwa mu ACSF ya okosijeni pa 32-34 ° C kwa maola 1-1.5 kuti alole kuchira. Panthawiyi, mawonekedwe a ionic, osmolarity, ndi mphamvu zowonjezera (monga shuga) za Artificial CSF ndizofunika kwambiri kuti zisungidwe kukhulupirika kwa minofu ndi mphamvu ya maselo.

Panthawi yojambulira ma electrophysiological, Artificial cerebrospinal fluid iyenera kuwonjezeredwa mosalekeza kuti pakhale malo okhazikika a extracellular. Izi zimatsimikizira kusinthasintha kwa pH, kutentha kwa ion, ndi kutentha, komanso kuchotsa zinyalala za metabolic, potsirizira pake zimathandizira kupeza deta yodalirika komanso yobwereketsa.

2. 2) Kuyesa Kwambiri Kwapadera Kwa Neuron Patch Clamp

Pakuwonongeka kwakukulu kwa neuronal, minofu ya muubongo nthawi zambiri imagayidwa ndi ma protease. Yankho la enzymatic limakonzedwa pamaziko a Artificial cerebrospinal fluid kuti mawonekedwe a nembanemba ndi ntchito ya ion njira zisungidwe pakagayidwe. Pambuyo pa chimbudzi ndi trituration yamakina, ma neuroni omwe amadzipatula amatsuka mobwerezabwereza ndi ACSF kuchotsa enzyme yotsalira ndikuyimitsidwanso mu ACSF kuti ipumule kapena kumamatira. Kukhazikika kwa osmolarity ndi pH ndizofunikira panthawi yonseyi kuti tipewe kuvulala kwa ma cell ndi apoptosis.

3. 3) Maselo Okhazikika ndi Ma cell Lines

Kwa muyeso wathunthu-kujambulira kwa cell ndi single-kujambulira kanjira, madzimadzi okhazikika amtundu wa cerebrospinal amatha kugwiritsidwa ntchito mwachindunji ngati njira yowonjezerapo kuti asungire kuthekera kwa nembanemba yopumula komanso momwe thupi limayendera njira za ion.

Posankha kudzipatula kwa ma ionic mafunde enaake, chimango cha Artificial CSF chimasinthidwa nthawi zambiri. Mwachitsanzo, pojambula mafunde a calcium, gawo la NaCl likhoza kusinthidwa ndi TEA-Cl kuti atseke njira za potaziyamu, kapena sodium ikhoza kuchotsedwa kuti itseke mafunde a sodium. Mukajambulitsa cholandirira cha NMDA-mafunde apakatikati, extracellular Mg2+ itha kusiyidwa kuti muchepetse block block. Nthawi zonse, zofunikira za physicochemical yankho ziyenera kukhala zokhazikika.

4. 4) Mayesero Ophatikizana Ogwira Ntchito

Artificial cerebrospinal fluid imagwiritsidwa ntchito pokonzekera mayankho ogwira ntchito a ion channel modulators ndi neuroactive compounds mu maphunziro a pharmacology, omwe amaikidwa kudzera mu perfusion system. Kupatuka kwa pH kapena kaphatikizidwe ka ionic kumatha kusintha kusungunuka kwa mankhwala, kukhazikika, kapena ngakhale kupangidwa kwa mapuloteni omwe akutsata, zomwe zimapangitsa kuti pakhale zotsatira zolakwika.

M'mayesero ophatikizika a optogenetic, zochitika za neuronal zimasinthidwa ndi kukondoweza kwa kuwala kenako ndikuwunikidwa ndi chigamba. Kuthirira mosalekeza ndi ACSF kumafunika kusunga maziko a electrophysiological state, kotero kuti kusintha komwe kumawoneka kungabwere chifukwa cha kulowererapo kwa optogenetic m'malo mosintha chilengedwe.

Malingaliro Aukadaulo ndi Zofunikira Zoyeserera

Pakuyesa kwa zigamba, zoyeserera zotsatirazi za Artificial Cerebrospinal Fluid-zogwirizana ziyenera kuyendetsedwa mosamalitsa kuti zitsimikizire kukhazikika kwa ma siginecha ndi kuberekanso.

Zonse pamodzi, zinthuzi zimatsimikizira kukhazikika kwa kujambula, kubwereza kwa deta, ndi kugwirizana kwachilengedwe kwa zotsatira.

Mapeto
Artificial cerebrospinal fluid ndi zambiri kuposa yankho lakumbuyo. Muukadaulo wa patch clamp, umagwira ntchito ngati maziko oyambira omwe amathandizira kuti minyewa ikhale yolimba, imateteza mawonekedwe a electrophysiological, ndikuwunikira magwiridwe antchito odalirika. Kukhathamiritsa mosamala kwa kapangidwe ka ACSF ndi miyeso yoyesera ndiye pakatikati pamayendedwe apamwamba aliwonse amtundu wa electrophysiology.

Kuti zithandizire izi, IPHASE imapereka zinthu zapamwamba - zapamwamba za ACSF zopangidwa mosamalitsa, zokhala ndi ma ionic okhazikika, osmolarity, ndi pH. Izi zimawonetsetsa kuti zambiri-ku-zosasinthika komanso magwiridwe antchito odalirika, kuthandiza ofufuza kupeza zotsatira zoyeserera zokhazikika komanso zobwerezabwereza.