Просте методичне пристосування для мікроін'єкторних маніпуляцій і вимірювань на електроморфологічному чипі при мікроінтерферометричному контролі інтерфейсних і мембранних процесів на діапазоні товщини від 50 до 10000 ангстрем під різними кутами

Автор(и)

  • O. V. Gradov Institute of Energetic Problems of Chemical Physics RAS named by V.L. Talroze, Russian Federation
  • F. A. Nasirov Institute of Energetic Problems of Chemical Physics RAS named by V.L. Talroze, Russian Federation
  • A. A. Skrynnik Institute of Energetic Problems of Chemical Physics RAS named by V.L. Talroze, Russian Federation
  • A. G. Jablokov Institute of Energetic Problems of Chemical Physics RAS named by V.L. Talroze, Russian Federation

DOI:

https://doi.org/10.26641/1997-9665.2017.4.7-17

Ключові слова:

мікроелектроди, мікроінтерферометр, мікроперфузія, патч-кламп, мікроманіпуляція, перфузія одиночних клітин, цитоелектрофізіологічний чіп

Анотація

Мікроманіпуляції, перфузії і вимірювання, що проводяться з використанням скляних мікроелектродів, заповнених, як правило, електролітом, є класичною технікою експериментально-морфологічних і мембранно-електрофізіологічних досліджень на рівні окремих клітин і мембранних поверхонь. Стандартний (ефективний) діаметр скляного мікроелектрода в кінцевій області становить від 500 нм до менш ніж 100 нм, що перешкоджає використанню стандартних оптичних мікроскопів для його спостереження, відповідно до оптичних критеріїв (критерій Релєя і т.п.), оскільки при діаметрі конуса менше 500 нм він губиться в інтерференційної облямівці. Мікропроцесорним програмуванням пуллера (мікрокузні), що забезпечує витягування і розрив, хоча і можна досягти в відомих режимах заданих форм і діаметра кінця мікропіпеток, цей результат не є в повній мірі контрольованим в силу вищевказаних обмежень. У зв'язку з цим необхідне створення пристроїв контролю кінцевого фрагмента мікропіпеток як при отриманні, так і при експлуатації (внутрішньоклітинному або екстрацелюлярному введенні) в штатному режимі. При цьому необхідно, щоб даний метод дозволяв візуалізувати на зображенні клітини з мікроелектродами в реальному часі процеси, що відбуваються між ними, в залежності від типу і стану електрода, що дозволить нівелювати артефакти, з частотою систематичної помилки, що виникають при неконтрольованій експлуатації кінця мікропіпеток після застосування різних способів заливки електроліту (капілярного по Тасакі; вакуумного заповнення; заповнення спиртом з подальшим витісненням спирту по еквівалентній об'ємній характеристиці електролітом; заливка легкоплавкими сплавами як альтернатива рідким електролітам, що полегшує введення контакту хлорсрібного дроту). Нами пропонується конфігурація установки, що вирішує всі вищевказані проблеми шляхом введення інтерферометричного пристрою для мікроскопічного контролю мікроелектродів і мікроманіпулятора або мікроперфузора, вперше для даного типу оптичних приладів комбінованого з інтерферометричною оптичною схемою.

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