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.
Что-то мне подсказывает, что я изобретаю велосипед, а все давно ясно и изобретено!
В общем, люди добрые, помогите, пожалуйста!!
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