Measurement of live bacteria by Nomarski interference microscopy and stereologic methods as tested with macroscopic rod-shaped models.
نویسندگان
چکیده
A new method is proposed to measure bacterial cells under growth conditions. Bacterial cells, suspended in their growth medium, were attached to a cover slip with poly-L-lysine. The cover slip was inverted and placed on a glass microscope slide. To prevent dehydration of the medium, the edges of the cover slip were sealed to the microscope slide with clear fingernail polish. The bacteria on the slide were then quickly photographed with a Leitz light microscope, using Nomarski optics. The photographic negatives were then projected at a standard distance through a lens system, and the projected images of the whole cells were outlined by hand onto graph paper. The profile images so transcribed onto the graph paper were in effect transverse sections of each of the cells. Using stereologic grid and point counting techniques, the area of the cell transverse section as well as the perimeter or circumference of the transverse section were estimated. Formulae were developed so that both the volume and surface area of the whole cell could be ascertained from these area and circumference measurements. Since the efficacy of any measurements of surface area and volume of microscopic rod-shaped bacterial cells could be questioned, macroscopic rod-shaped models were used to test the theory and formulae and to compare this method with other commonly used cell-sizing techniques. This technique could be used in any study of bacterial cell size or changes in cell size (e.g., osmotic shifts).
منابع مشابه
Structural periodicities observed in mammalian rod outer segments with Nomarski optics.
Structural periodicities have been observed in isolated rod outer segments of several mammalian species using differential interference contrast (Nomarski) optics. The spacing among the observed structural inhomogeneities is somewhat variable. If these measured spacings are corrected for shrinkage, estimated by comparing ROS widths in unfixed cells to those in light microscopic autoradiographs,...
متن کاملThree-Dimensional Superresolution Imaging of the FtsZ Ring during Cell Division of the Cyanobacterium Prochlorococcus
Superresolution imaging has revealed subcellular structures and protein interactions in many organisms. However, superresolution microscopy with lateral resolution better than 100 nm has not been achieved in photosynthetic cells due to the interference of a high-autofluorescence background. Here, we developed a photobleaching method to effectively reduce the autofluorescence of cyanobacterial a...
متن کاملGreener Synthesis of Rod Shaped Zinc Oxide NPs Using Lilium ledebourii Tuber and Evaluation of Their Leishmanicidal Activity
Background: NPs (NPs) with unique chemical and physical properties can be used for therapeutic purposes because of their strong antimicrobial activates. NPs have been used as an antimicrobial agents to inhibit microbial growth.Objectives: In view of the strong antimicrobial activity of NPs, the biogenic synthesis and leishmanicidal activity of rod-shaped ...
متن کاملInvestigation of cytotoxicity properties of zinc oxide nanoparticles in spherical and rod shaped on leukemia cells
In this study, we reported a method to associate doxorubicin drug on folic acid functionalized SiO2/ZnO nanoparticles (NPs) in rod and spherical shapes. The clinical usage of the anticancer drug, doxorubicin (DOX), is limited by severe side effects and cell resistance. Targeted drug delivery by binding DOX to nanoparticles could overcome these limitations. The surface functionalization of the Z...
متن کاملMicrobial diversity within early-stage cultured Panulirus ornatus phyllosomas.
A thorough understanding of the microorganisms and pathogens associated with the larval stage of the tropical ornate rock lobster, Panulirus ornatus, is required to overcome disease outbreaks that currently block aquaculture attempts. This study used microscopy in addition to culture and molecularly based microbiological techniques to characterize the bacterial community associated with culture...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Applied and environmental microbiology
دوره 54 1 شماره
صفحات -
تاریخ انتشار 1988