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A Raman Flow Cytometer: An Innovative Microfluidic Approach for Continuous Label-Free Analysis of Cells via Raman Spectroscopy

機譯:拉曼流式細胞儀:一種創(chuàng)新的微流控方法,用于通過拉曼光譜連續(xù)無標簽分析細胞

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摘要

In this work a Raman flow cytometer is presented.ududIt is a whole new microfluidic device that takes advantage of basic principles of Raman spectroscopy and fluorescent flow cytometry mixed together in a system of particularly shaped channels. These are indeed composed by specific shape and sizes – thanks to which cells can flow one-by-one – and a trap by means of which cells are trapped in order to perform Raman analysis on single ones in a constant and passive way. In this sense the microfluidic device promotes a fast method to look for single cells in a whole multicellular sample.ududIt is a label-free analysis and this means that, on the contrary of what happens with fluorescent flow cytometry, the sample does not need to undergo any particular time-consuming pretreatment before being analyzed. Moreover it gives a complete information about the biochemical content of the sample thanks to the involvement of Raman spectroscopy as method of analysis.ududMany thought about a device like this, but eventually it is the first one being designed, fabricated and tested. The materials involved in the production of the Raman flow cytometer are chosen wisely. In particular the chip – the most important component of the device – is multilayered, being composed by a slide of calcium fluoride (which gives a negligible signal in Raman analyses), a photosensitive resist containing a pattern with channels and another slide of calcium fluoride in order for the channels to be sealed on both sides. The chip is, in turn, connected to gaskets and external frames. Several fabrication processes are followed to ultimately get the complete Raman flow cytometer and experiments on red blood cells demonstrate its validity in this field.
機譯:在這項工作中,將介紹拉曼流式細胞儀。 ud ud這是一種全新的微流控設備,它利用拉曼光譜學和熒光流式細胞儀的基本原理混合在一起,形成了特殊形狀的通道。這些的確是由特定的形狀和大小組成的-歸因于此,細胞可以一個一個地流動-并通過一個陷阱捕獲細胞,以便以恒定和被動的方式對單個細胞進行拉曼分析。從這個意義上說,微流體裝置促進了一種快速的方法來尋找整個多細胞樣品中的單細胞。 ud ud這是一種無標記的分析,這意味著與熒光流式細胞術相反,該樣品可以在進行分析之前,不需要進行任何特定的耗時的預處理。此外,由于拉曼光譜法作為分析方法,它提供了有關樣品生化成分的完整信息。 ud ud許多人想到了這樣的設備,但最終它是第一個被設計,制造和測試的設備。拉曼流式細胞儀生產中涉及的材料是明智選擇的。尤其是該芯片(該設備最重要的組件)是多層的,由氟化鈣片(在拉曼分析中給出的信號可忽略不計),光敏抗蝕劑組成,該光刻膠包含帶有通道的圖案,而另一片氟化鈣片包含在其中。以便在兩側密封通道。芯片又連接到墊圈和外部框架。遵循幾個制造過程以最終獲得完整的拉曼流式細胞儀,對紅細胞的實驗證明了其在該領域的有效性。

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    De Grazia Antonio;

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  • 年度 2015
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