Mikrostrukturierten Durchflusszytometer

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Inge Bergmann
vor 7 Jahren
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1 Hydrodynamische Fokussierung in Mikrostrukturierten Durchflusszytometer Hydrodynamische Fokussierung Patentstruktur Neue Strukturen Was, Wozu TU Berlin / PTB RWTH Aachen, Micronit Microfluidics BV

Hydrodynamic focussing Flow cytometry: hydrodynamic focussing: Sample stream ensheathed by fluid and mechanically focused by a funnel or the like Microfluidics: sheath flow focussing In microfluidics

2 Hydrodynamic focussing Flow cytometry: hydrodynamic focussing: Sample stream ensheathed by fluid and mechanically focused by a funnel or the like Microfluidics: sheath flow focussing In microfluidics 'sheath flow focussing' can also mean focussing of particle path using patterned channels (e.g. using chevrons, Dean vortices, constriction-expansion) Flow cytometry Microfluidics Meaning 1-D focussing 2-D focussing Sample stream focussed in 1 lateral dimension 2-D focussing 3-D focussing Sample stream focussed in 2 lateral dimensions

3 Hydrodynamic focussing Why? Exactly position inside sheath flow: Avoid adhesion on channel walls High signal stability (e.g. sample position in laser beam profile) optical flow cytometry mixture ultra rainbow beads 8 peaks calibration

4 Hydrodynamic focussing Why? Exactly position inside sheath flow: Avoid adhesion on channel walls High signal stability (e.g. sample position in laser beam profile) Ein neuartiges 3-Dmikrostrukturiertes Durchflusszytometer, TU Berlin 2007 optical flow cytometry mixture ultra rainbow beads 8 peaks calibration Mikrofluidische Sensoren in der Durchflusszytometrie zur kombinierten optischelektrischen Analyse an Blutzellen, RWTH Aachen 2016

5 Hydrodynamic focussing Why? Exactly position inside sheath flow: Avoid adhesion on channel walls High signal stability (e.g. sample position in laser beam profile) Ein neuartiges 3-Dmikrostrukturiertes Durchflusszytometer, TU Berlin 2007 optical flow cytometry mixture ultra rainbow beads 8 peaks calibration Mikrofluidische Sensoren in der Durchflusszytometrie zur kombinierten optischelektrischen Analyse an Blutzellen, RWTH Aachen 2016

6 Hydrodynamic focussing µfcm-pc Patent Janko Theissen et al. (TUB, PTB) measurement channel focussing Spin focussing sheath fluid sample fluid

7 Hydrodynamic focussing µfcm-pc Patent Janko Theissen et al. (TUB, PTB) Microfluidic chip works for 2D (3D) focussing Chip produced by hot embossing: plastic material only! Kummrow A, Theisen J, Frankowski M, Tuchscheerer A, Yildirim H, Brattke K, Schmidt M, Neukammer J, Lab Chip 9 (2009),

8 Hydrodynamic focussing µfcm Need to have better material: virus Frankowski M, Bock N, Kummrow A, Schädel-Ebner S, Schmidt M, Tuchscheerer A, Neukammer J, Cytometry A 79A (2011), Cells Technology (PDMS) hot embossing soft lithography Frankowski M, Simon P, Bock N, El-Hasni A, Schnakenberg U, Neukammer J, Engineering in Life Sciences (2015) lithography & glass etching

9 Hydrodynamic focussing µfcm-glass Microfluidic chip has better optical quality But: no 2D (3D) hydrodynamic focussing 1D focussing only Reason unclear. Design flaw? Manufacturing issue? µfcm-pc µfcm-glass top sheath sample TMMF laminate bottom sheath

Summary: Microfluidic flow cytometer chip Measurands of flow cytometry require specific optical properties of the construction material (low light scatter, low background fluorescence) Construction

10 Summary: Microfluidic flow cytometer chip Measurands of flow cytometry require specific optical properties of the construction material (low light scatter, low background fluorescence) Construction material requires specific micro-manufacturing processes (structuring 2D/3D, electrical/optical wiring, mounting) Micro-fabrication process requires specific fluidic design Reliable modulation for flow of sample suspensions in microfluidic flow cytometers is missing Example: filamentation of sample stream upon hydrodynamic focusing

11 Physikalisch-Technische Bundesanstalt Braunschweig und Berlin Abbestr Berlin Andreas Kummrow Working Group 8.32: Flow Cytometry and Microscopy Phone: ++49-(0)

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