Sub-Attomole Molecule Detection in a Single Biological Cell in-vitro by Thermal Lens Microscopy

Harada, Masaaki; Shibata, Masashi; Kitamori, Takehiko; Sawada, Tsuguo

doi:10.2116/analsci.15.647

Cited by 38 publications

(14 citation statements)

References 16 publications

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“…11,12 To overcome this problem, we have developed a photothermal microscope based on the thermal-lens effect, the thermal-lens microscope (TLM), and have reported that it can be applied to the ultra-sensitive detection of non-fluorescent substances. [13][14][15][16][17][18][19] Furthermore, we found that the thermal-lens signal measured in the micro-channel was more stable than that measured in much wider space, which showed that TLM is very suitable as a micro-space detection method in an integrated chemistry laboratory, ICL. 13 Urinary levels of AA, especially following a load test, can be helpful in the clinical diagnosis of scurvy.…”

Section: Introductionmentioning

confidence: 80%

Integrated FIA for the Determination of Ascorbic Acid and Dehydroascorbic Acid in a Microfabricated Glass-Channel by Thermal-Lens Microscopy

et al. 2000

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An on-chip chemical micro-determination device, integrating the FIA system, was applied to L-ascorbic acid (AA) and dehydroascorbic acid (DHAA) determination. Microchannels having a 250×100 µm cross-section were fabricated in a quarts glass chip as a component in an integrated FIA system. They were put to use for flow, mixing, reaction, and detection. The reaction system was a coupled redox-complexation reaction between AA and a 1,10-phenantroline-Fe(III) mixture. Sample solutions were introduced into micro-channels by micro-syringes, the flow rate was controlled by a micro-syringe pump, and the reagents, were mixed by molecular diffusion. A photothermal microscope was used for the ultra-sensitive detection of the non-fluorescent reaction product (ferroin). For DHAA determination, dithiothreithol was used as a reducing agent and the total AA was determined. The DHAA quantity was calculated by subtraction. This method allows for the determination of AA with a linear range of up to 40 µM. The RSD for 25 µM AA (n=5) is 1%, and the limit of detection (LOD) is 0.1 µM. The procedure was successfully applied to determining the AA and DHAA content of urine and AA in vitamin C tablets. The results agree well with those obtained by reference methods.

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Section: Introductionmentioning

confidence: 80%

Integrated FIA for the Determination of Ascorbic Acid and Dehydroascorbic Acid in a Microfabricated Glass-Channel by Thermal-Lens Microscopy

et al. 2000

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“…TLM has been applied to other miniaturized systems and demonstrated to be a suitable detector for them. [22][23][24] Then we used TLM to detect the DNA fragments separated in the slab gel with the intent of reducing band-broadening from the detector itself.…”

Section: Resultsmentioning

confidence: 99%

Fast Slab Gel Electrophoretic Separation of DNA Fragments with a Short Migration Distance Using Thermal Lens Microscope

et al. 1999

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We demonstrated fast separation of DNA fragments in a short slab gel through the minimization of band-broadening. A discontinuous buffer system was used to reduce band-broadening from sample introduction, and a high spatial resolution thermal lens microscope (TLM) was used to reduce such an effect from the detector. Bands of DNA fragments were sharpened into 100 µm, which was about 1/10 of the bandwidths in conventional continuous slab gel electrophoresis. The 100 base pair ladder DNA sample, which ranges from 100 bp to 2000 bp, was completely separated within 15 min with a migration distance of 18 mm and was detected by TLM after silver staining. These results suggested the possibility of fast separation of DNA fragments in a miniaturized discontinuous slab gel. As well as the inherent properties of parallel separation of slab gel, high throughput separation can be obtained, which is significant for DNA analyses such as DNA sequencing, genetic diagnosis and forensic identification analyses.

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“…As to sensitivity, TLM can detect a single solute molecule in the detection volume, where the limit of detection (LOD) is estimated by a time-averaged value [44,45]. With regard to the high applicability and high spatial resolution, TLM could be applied to determination of target solute in free solution in an optical cell [46], DNA detection in polymer matrix [47], dye molecule determination in a single cell [48], and detection of a nm-sized single particle in solution [49]. LODs of TLM for various samples are summarized in Table 1.…”

Section: Thermal Lens Microscopementioning

confidence: 99%

Chemical processing on microchips for analysis, synthesis, and bioassay

et al. 2003

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Sub-Attomole Molecule Detection in a Single Biological Cell in-vitro by Thermal Lens Microscopy

Cited by 38 publications

References 16 publications

Integrated FIA for the Determination of Ascorbic Acid and Dehydroascorbic Acid in a Microfabricated Glass-Channel by Thermal-Lens Microscopy

Integrated FIA for the Determination of Ascorbic Acid and Dehydroascorbic Acid in a Microfabricated Glass-Channel by Thermal-Lens Microscopy

Fast Slab Gel Electrophoretic Separation of DNA Fragments with a Short Migration Distance Using Thermal Lens Microscope

Chemical processing on microchips for analysis, synthesis, and bioassay

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