Quantitative analysis of very small quantities of cultured cells

In this paper, methods of quantitative elemental analysis of very small quantity of cultured cells were developed. First of all, an internal-standard method for the solution containing cells whose density is more than 1 × 106 cells/mL was established and then a standard-free method for cultured cells was developed. It was confirmed that the method allows us to quantitatively analyze more than 25 elements in the samples containing only 20,000 cells. Also, the methods for removing cultured cells from a flask were examined in order to improve accuracy and sensitivity of analysis since the use of trypsin and PBS sometimes brings a large amount of sodium, phosphorus and potassium, which have direct effect upon accuracy of analysis based on the standard-free method. It was found that the method of removing cells with a scraper without using trypsin and PBS is the best manner. Also, the effects of using thinner backing materials were examined in order to improve sensitivity of analyses. It is expected that accurate analysis of samples containing nearly two thousand cells is possible on the basis of the standard-free method when using a thinner backing material.

[1]  K. Sera,et al.  ANALYSIS OF THE LETHAL EFFECTS OF LOW-DOSE RADIATION ON GLIOMA CELLS: RELATIONSHIP OF THE BYSTANDER EFFECT AND METAL ELEMENTS , 2011 .

[2]  Ahmad Faried,et al.  Quantitative analysis of cisplatin sensitivity of human esophageal squamous cancer cell lines using in‐air micro‐PIXE , 2010, Cancer science.

[3]  D. Brenner,et al.  Tumor Necrosis Factor Alpha-Induced Interleukin-8 Production via NF-κB and Phosphatidylinositol 3-Kinase/Akt Pathways Inhibits Cell Apoptosis in Human Hepatocytes , 2002, Infection and Immunity.

[4]  S. Futatsugawa,et al.  Quantitative analysis of untreated hair samples for monitoring human exposure to heavy metals , 2002 .

[5]  S. Futatsugawa,et al.  Quantitative analysis of untreated bio-samples , 1999 .

[6]  K. Sera,et al.  Personal computer aided data handling and analysis for PIXE , 1996 .

[7]  Werner Brandt,et al.  Energy-loss effect in inner-shell Coulomb ionization by heavy charged particles , 1981 .

[8]  E. Weisburger,et al.  Ratliver cells in culture: Effect of storage, long-term culture, and transformation on some enzyme levels , 1976, In Vitro.

[9]  R. Rajesh,et al.  Anticancer effects of brominated indole alkaloid Eudistomin H from marine ascidian Eudistoma viride against cervical cancer cells (HeLa). , 2015, Anticancer research.

[10]  K. Sera,et al.  Quantitative analysis of small bio-samples of nearly 1 μg , 2014 .

[11]  G. Wakabayashi,et al.  QUANTITATIVE ANALYSIS OF VERY SMALL QUANTITY OF ORGANS TAKEN FROM PATIENTS AND EXPERIMENTAL ANIMALS: STANDARD-FREE METHOD FOR ORGAN SAMPLES , 2008 .

[12]  K. Sera,et al.  STANDARD-FREE METHOD FOR BODY FLUID SAMPLES SUCH AS SALIVA, SWEAT, SNIVAL AND TEAR , 2005 .

[13]  M. Carrière,et al.  HEAVY METALS TOXICITY AFTER ACUTE EXPOSURE OF CULTURED RENAL CELLS: INTRACELLULAR ACCUMULATION AND REPARTITION , 2005 .

[14]  S. Matsuyama,et al.  THE ELEMENTAL ANALYSIS OF IgE-SENSITIZED RBL-2H3 CELLS USING IN-AIR MICRO-PIXE , 2004 .

[15]  K. Sera,et al.  APPLICATION OF A STANDARD-FREE METHOD TO QUANTITATIVE ANALYSIS OF URINE SAMPLES , 2001 .

[16]  K. Sera,et al.  DIFFERENTIAL RESPONSES OF COPPER(I) AND COPPER(II) ON LIVER COPPER ACCUMULATION IN THE LONG-EVANS CINNAMON (LEC) RATS STUDIED BY PIXE DETERMINATION , 2000 .

[17]  K. Sera,et al.  QUANTITATIVE ANALYSIS OF BIO-MEDICAL SAMPLES OF VERY SMALL QUANTITIES BY THE STANDARD-FREE METHOD , 1997 .

[18]  K. Sera,et al.  STANDARD-FREE METHOD OF QUANTITATIVE ANALYSIS FOR BIO-SAMPLES , 1996 .

[19]  Ping Huang,et al.  STUDY OF TRACE ELEMENTS IN CELLS USING SPM, SR-XRM AND PIXE , 1995 .

[20]  K. Sera,et al.  PRESENT STATUS OF NMCC AND SAMPLE PREPARATION METHOD FOR BIO-SAMPLES , 1993 .

[21]  K. Sera,et al.  Bio-PIXE AT THE TAKIZAWA FACILITY (Bio-PIXE WITH A BABY CYCLOTRON) , 1992 .