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Визуализация липофусцина в конфокальном и эпи-флуоресцентном микроскопе.

mosmasha, 30.11.2014 20:37
Уважаемые коллеги! Подскажите, пожалуйста, как правильно работать с аутофлуоресценцией липофусцина.

Я работаю на нейронах гиппокампа - на 440-460 нм при возбуждении 405нм лазером (конфокал) светится все! Оно и понятно- это НАДН, но мне нужен липофусцин. Со временем, когда клетки уже в буфере, флуоресценция НАДН затухает, потому как окисляется весь эндогенный НАДН.
А нельзя ли этот процесс ускорить? Затушить всю флуоресценцию НАДН и быть точно уверенным, что светит только липофусцин.
В литературе предлагают смотреть на 530нм при возбуждении 488нм лазером, но там же флавиновая полоса, причем флавины наоборот начинают светить когда окисляются. confused.gif


Что-то мне подсказывает, что я изобретаю велосипед, wall.gif а все давно ясно и изобретено!

В общем, люди добрые, помогите, пожалуйста!!
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