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Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.

Miyake M, Nakai Y, Anai S, Tatsumi Y, Kuwada M, Onishi S, Chihara Y, Tanaka N, Hirao Y, Fujimoto K - Cancer Sci. (2014)

Bottom Line: The overall sensitivities of conventional cytology, BTA, NMP22, fluorescence cytology, fluorescent spectrophotometric assay and CFAU assay were 48%, 33%, 40%, 86%, 86% and 87%, respectively.Three different ALA-based assays showed high sensitivity and specificity.Development of a rapid and automated device for ALA-based photodynamic assay is necessary to avoid the variability induced by troublesome steps and low stability of specimens.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.

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Related in: MedlinePlus

Quantitative detection of protoporphyrin IX using a fluorescence spectrophotometer. Fluorescent spectra of representative cases are shown. Red curves are spectra of samples treated with ALA, and blue curves are spectra of non-ALA control. Fluorescence intensities at 635 nm are compared between ALA-treated and untreated cells. The assay was run in triplicate. Both cases with bladder cancer are considered positive for the fluorescence spectrophotometric assay. a.u., arbitrary unit; LG, low grade; HG, high grade. †The intensity value of ALA-treated samples is higher than those of ALA-untreated controls.
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fig03: Quantitative detection of protoporphyrin IX using a fluorescence spectrophotometer. Fluorescent spectra of representative cases are shown. Red curves are spectra of samples treated with ALA, and blue curves are spectra of non-ALA control. Fluorescence intensities at 635 nm are compared between ALA-treated and untreated cells. The assay was run in triplicate. Both cases with bladder cancer are considered positive for the fluorescence spectrophotometric assay. a.u., arbitrary unit; LG, low grade; HG, high grade. †The intensity value of ALA-treated samples is higher than those of ALA-untreated controls.

Mentions: We used 50 mL of voided urine for each of the ALA-treated sediments and ALA-untreated sediments. After the treatment with ALA in serum-free media, urine sediments were obtained by centrifugation and resuspended in 500 μL of PBS. The resuspended sediments in PBS were transferred in triplicate (100 μL each) to a flat-bottom transparent 96-well plate. In each urine sample, ALA-treated sediments and untreated sediments were subjected to the spectrophotometric assay. Fluorescence emission spectra were recorded using a microplate spectrophotometer (Infinite 200M PRO; Tecan, Männedorf, Switzerland) equipped with i-control version 1.8 software. The excitation wavelength was 400 nm, and the photomultiplier tube voltage was fixed. A series of spectra obtained from each specimen (Fig.3) and the fluorescence intensity at 635 nm was measured. The values of ALA-treated sediments and those of the ALA-untreated control were compared. If all of the three intensity values of ALA-treated samples were higher than any values of ALA-untreated controls, the sample was considered positive for the spectrophotometric assay.


Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.

Miyake M, Nakai Y, Anai S, Tatsumi Y, Kuwada M, Onishi S, Chihara Y, Tanaka N, Hirao Y, Fujimoto K - Cancer Sci. (2014)

Quantitative detection of protoporphyrin IX using a fluorescence spectrophotometer. Fluorescent spectra of representative cases are shown. Red curves are spectra of samples treated with ALA, and blue curves are spectra of non-ALA control. Fluorescence intensities at 635 nm are compared between ALA-treated and untreated cells. The assay was run in triplicate. Both cases with bladder cancer are considered positive for the fluorescence spectrophotometric assay. a.u., arbitrary unit; LG, low grade; HG, high grade. †The intensity value of ALA-treated samples is higher than those of ALA-untreated controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317833&req=5

fig03: Quantitative detection of protoporphyrin IX using a fluorescence spectrophotometer. Fluorescent spectra of representative cases are shown. Red curves are spectra of samples treated with ALA, and blue curves are spectra of non-ALA control. Fluorescence intensities at 635 nm are compared between ALA-treated and untreated cells. The assay was run in triplicate. Both cases with bladder cancer are considered positive for the fluorescence spectrophotometric assay. a.u., arbitrary unit; LG, low grade; HG, high grade. †The intensity value of ALA-treated samples is higher than those of ALA-untreated controls.
Mentions: We used 50 mL of voided urine for each of the ALA-treated sediments and ALA-untreated sediments. After the treatment with ALA in serum-free media, urine sediments were obtained by centrifugation and resuspended in 500 μL of PBS. The resuspended sediments in PBS were transferred in triplicate (100 μL each) to a flat-bottom transparent 96-well plate. In each urine sample, ALA-treated sediments and untreated sediments were subjected to the spectrophotometric assay. Fluorescence emission spectra were recorded using a microplate spectrophotometer (Infinite 200M PRO; Tecan, Männedorf, Switzerland) equipped with i-control version 1.8 software. The excitation wavelength was 400 nm, and the photomultiplier tube voltage was fixed. A series of spectra obtained from each specimen (Fig.3) and the fluorescence intensity at 635 nm was measured. The values of ALA-treated sediments and those of the ALA-untreated control were compared. If all of the three intensity values of ALA-treated samples were higher than any values of ALA-untreated controls, the sample was considered positive for the spectrophotometric assay.

Bottom Line: The overall sensitivities of conventional cytology, BTA, NMP22, fluorescence cytology, fluorescent spectrophotometric assay and CFAU assay were 48%, 33%, 40%, 86%, 86% and 87%, respectively.Three different ALA-based assays showed high sensitivity and specificity.Development of a rapid and automated device for ALA-based photodynamic assay is necessary to avoid the variability induced by troublesome steps and low stability of specimens.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.

Show MeSH
Related in: MedlinePlus