4 In Situ Nick End-Labeling: Electron Microscopical
45
tion products in the conventional ISEL technique or immunogold complexes in the ISELITEM technique. Methyl green has
been known to stain double-stranded DNA (Burres et al.
1993; Melnick and Pickering 1988) and is weakly auto fluorescent
(Rothbarth et aI1976). The ISEL using eLM technique combined
with methyl green staining enables the simultaneous localization
of both 3' -OH DNA ends and double-stranded DNA in apoptotic
cells and also, the comparison of their concentrations (Ito and
Otsuki 1998).
References
Ansari B, Coates PJ, Greenstein BD, Hall PA (1993) In situ end-labeling detects DNA strand breaks in apoptosis and other physiological and pathological states. J Pathol 170: 1-8
Arends MJ, Morris RG, Wyllie AH (1990) Apoptosis: the role of the endonuclease. Am J Pathol 136:593-608
Burres NS, Frigo A, Rasmussen RR, McAlpine JB (1992) A colorimetric microassay for the detection of agents that interact with DNA. J Nat Prod
55:1582-1587
Gavrieli Y, Sherman Y, Ben-SAsson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell BioI 119:493-501
Herrmann M, Lorenz HM, VollR, Griinke M, Woith W, KaldenJR (1994) A
rapid and simple method for the isolation of apoptotic DNA fragments.
Nucleic Acids Research 22:5506-5507
Inoki C, Ito Y, Yamashita H, Ueki K, Fukuda Y, Ninomiya E, Nakamura K,
Hayashi R, Ueki M, Otsuki Y (1997) Image analysis and ultrastructural
detection of DNA strand breaks in human endometrium by in situ endlabeling techniques. J Histotechno120:321-328
Ito Y, Otsuki Y (1998) Localization of apoptotic cells in the human epidermis by an in situ DNA nick end labeling method using confocal reflectant
laser microscopy. J Histochem Cytochem (in press)
Kerr JFR, Wyllie AH, Currie AR(1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer
26:239-257
Melnick J, Pickering M (1988) The mechanism of the drug induced partial
displacement of methyl green from DNA. Biochem Int 16:69-75
Rothbarth PH, Hendriks-Sturkenboom I, Ploem JS (1976) Identification of
monocytes in suspensions of mononuclear cells. Blood 48:139-147
Thiry M (1991) In situ nick translation at the electron microscopic level: a
tool for studying the location of DNase-sensitive regions within the cell.
J Histochem Cytochem 39:871-874
45
tion products in the conventional ISEL technique or immunogold complexes in the ISELITEM technique. Methyl green has
been known to stain double-stranded DNA (Burres et al.
1993; Melnick and Pickering 1988) and is weakly auto fluorescent
(Rothbarth et aI1976). The ISEL using eLM technique combined
with methyl green staining enables the simultaneous localization
of both 3' -OH DNA ends and double-stranded DNA in apoptotic
cells and also, the comparison of their concentrations (Ito and
Otsuki 1998).
References
Ansari B, Coates PJ, Greenstein BD, Hall PA (1993) In situ end-labeling detects DNA strand breaks in apoptosis and other physiological and pathological states. J Pathol 170: 1-8
Arends MJ, Morris RG, Wyllie AH (1990) Apoptosis: the role of the endonuclease. Am J Pathol 136:593-608
Burres NS, Frigo A, Rasmussen RR, McAlpine JB (1992) A colorimetric microassay for the detection of agents that interact with DNA. J Nat Prod
55:1582-1587
Gavrieli Y, Sherman Y, Ben-SAsson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell BioI 119:493-501
Herrmann M, Lorenz HM, VollR, Griinke M, Woith W, KaldenJR (1994) A
rapid and simple method for the isolation of apoptotic DNA fragments.
Nucleic Acids Research 22:5506-5507
Inoki C, Ito Y, Yamashita H, Ueki K, Fukuda Y, Ninomiya E, Nakamura K,
Hayashi R, Ueki M, Otsuki Y (1997) Image analysis and ultrastructural
detection of DNA strand breaks in human endometrium by in situ endlabeling techniques. J Histotechno120:321-328
Ito Y, Otsuki Y (1998) Localization of apoptotic cells in the human epidermis by an in situ DNA nick end labeling method using confocal reflectant
laser microscopy. J Histochem Cytochem (in press)
Kerr JFR, Wyllie AH, Currie AR(1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer
26:239-257
Melnick J, Pickering M (1988) The mechanism of the drug induced partial
displacement of methyl green from DNA. Biochem Int 16:69-75
Rothbarth PH, Hendriks-Sturkenboom I, Ploem JS (1976) Identification of
monocytes in suspensions of mononuclear cells. Blood 48:139-147
Thiry M (1991) In situ nick translation at the electron microscopic level: a
tool for studying the location of DNase-sensitive regions within the cell.
J Histochem Cytochem 39:871-874