Microfluidic channel-assisted screening of hematopoietic malignancies

Farah Mughal, Sara J. Baldock, Ehsan Ghayoor Karimiani, Nick Telford, Nicholas J. Goddard, Philip J R Day

Research output: Contribution to journalArticlepeer-review

Abstract

A simple microfluidic fluorescence in situ hybridization (FISH) device allowing accurate analysis of interphase nuclei in 1 hr in narrow channels is presented. Photolithography and fluorosilicic acid etching were used to fabricate microfluidic channels (referred to as FISHing lines) that allowed analysis of 10 samples on a glass microscope slide 0.2 μl of sample volume was used to fill a micro-channel, which resembled a 250-fold reduction compared to conventional FISH. FISH signals were comparable to conventional FISH, with 50-fold less probe consumption and 10-fold less time. Cells were immobilized in single file in channels just exceeding the diameter of the cells, and were used for minimal residual disease (MRD) analysis. To test the micro-channels for application in FISH, MRD was simulated by mixing K562 cells (an established chronic myeloid leukemia cell line) carrying the BCR/ABL fusion gene across 1:1 to 1:1,000 Jurkat cells (an established acute lymphoblastic leukemia cell line). The limit of detection was seen to be 1:100 cells and 1:1,000 cells for FISHing lines and conventional FISH, respectively; however, the conventional method seemed to over-score the presence of K562 cells. This may in part be attributed to FISHing lines practically eliminating the chance of duplicate screening of cells and hastened the time of screening, enhancing scoring of all cells within the channels. This was compared to 1 in 500 cells on the slide being analyzed with the conventional FISH. © 2013 Wiley Periodicals, Inc.
Original languageEnglish
Pages (from-to)255-263
Number of pages8
JournalGenes Chromosomes and Cancer
Volume53
Issue number3
Early online date16 Dec 2013
DOIs
Publication statusPublished - Mar 2014

Keywords

  • Gene Role: ANT (Analyte), BSU (Biological study, unclassified), ANST (Analytical study), BIOL (Biological study) (ABL fusion with BCR
  • microfluidic channel-assisted screening of hematopoietic malignancies)
  • Gene Role: ANT (Analyte), BSU (Biological study, unclassified), ANST (Analytical study), BIOL (Biological study) (BCR, fusion with ABL
  • Animal cell line (human Jurkat cell line
  • Animal cell line (human K-562 cell line
  • Nucleic acid hybridization (in situ, fluorescence
  • Acute lymphocytic leukemia
  • Animal tissue culture
  • Biosensors
  • Chronic myeloid leukemia
  • Electronic device fabrication
  • Etching
  • Fluorescence
  • Fluorescent indicators
  • Glass substrates
  • Human
  • Microfluidic devices
  • Nucleic acid hybridization
  • Photolithography (microfluidic channel-assisted screening of hematopoietic malignancies)
  • Chimeric gene Role: ANT (Analyte), BSU (Biological study, unclassified), ANST (Analytical study), BIOL (Biological study) (microfluidic channel-assisted screening of hematopoietic malignancies)
  • Probes Role: ARG (Analytical reagent use), BUU (Biological use, unclassified), PEP (Physical, engineering or chemical process), TEM (Technical or engineered material use), ANST (Analytical study), BIOL (Biological study), PROC (Process), USES (Uses) (mi
  • microfluidic device channel FISH chronic myeloid acute lymphoblastic leukemia

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