Fluorescent Protein-Based Biosensors and Their Clinical Applications - Progress in Molecular Biology and Translational Science - page 336

Table 8.2 Specifications of Flicyme
Light source 407 or 445 nm laser diode (60 mW)

Detection parameter
and detector

Scattering light, 2 ch: forward scatter, photodiode;
side scatter, PMT
Fluorescence intensity and lifetime, 3 ch: PMT

Detection fluorescence
wavelength

Ch1: 482/35 nm (e.g., CFP or AG)
Ch2: 542/30 nm (e.g., YFP) or 579/34 nm
(e.g., KO)
Ch3: 650-long pass (e.g., Keima)

Sample flow rate 6 m/s

Sample flow volume
(

Lo: 40, mid: 80, hi: 160

l min -1 ) m

Maximum acquisition rate 10,000 events/s

Modulation frequency 28 MHz

Size (mm) 500 ( W ) 675 ( D ) 700 ( H )

PMT, photomultiplier; AG, Azami Green; KO, Kusabira Orange.

including initial screening of FRET constructs (to select one with greater
gains), drug screening in which comparison of FRET efficiencies among
samples is required, and detection of a small subpopulation within a hetero-
geneous population via observation of a large number of cells (see below).

7. CLINICAL APPLICATION OF FRET
In the latter part of this review, we would like to introduce a novel di-
agnostic method based on bioimaging technology and fluorescent proteins,
using chronic myeloid leukemia (CML) as a model. CML, a hematological
malignancy involving the transformation of hematopoietic stem cells in the
bone marrow, is characterized by the formation of an abnormal chromosome
(Philadelphia chromosome, Ph1) and the expression of its transcript BCR-
ABL. 108 About 8000 people in Japan and 63,000 worldwide, representing
about one-fifth to one-fourth of all leukemia patients, suffer from this disease.
BCR-ABL encodes a constitutively active tyrosine kinase and causatively
contributes to malignant transformation of the leukemia cells by activating a
range of signaling pathways through tyrosine phosphorylation of its substrates,
including CrkL and signal transducer and activator of transcription. 109,110