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Related Concept Videos

Spectrophotometry: Introduction01:16

Spectrophotometry: Introduction

2.9K
Spectrophotometry is the quantitative measurement of the absorption, reflection, diffraction, or transmission of electromagnetic radiation through a material as a function of the intensity and wavelength of the radiation. A spectrophotometer is a device used to measure the change in the radiation intensity caused by its interaction with the material.
The essential components of a spectrophotometer include a source of electromagnetic radiation, a slot for placing a material to be analyzed, and a...
2.9K
UV–Vis Spectrometers01:14

UV–Vis Spectrometers

1.3K
The absorbance of UV and visible (UV–visible) radiations is measured using a UV–visible spectrophotometer. Deuterium lamps, which emit UV radiation, and tungsten lamps, which produce radiation in the visible region, are used as light sources in UV–visible spectrophotometers. A monochromator or prism is used for diffraction grating, i.e., to split the incoming radiation into different wavelengths. A system of slits is used to focus the desired wavelength on the sample cell.
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Microbial Growth Measurement: Indirect Methods01:27

Microbial Growth Measurement: Indirect Methods

Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
UV–Vis Spectroscopy: Beer–Lambert Law01:09

UV–Vis Spectroscopy: Beer–Lambert Law

2.1K
The Beer-Lambert law describes the relationship between absorbance and concentration, which combines the principles established by scientists Johann Heinrich Lambert and August Beer. Lambert's law states that when light passes through a medium, the loss in intensity is directly proportional to the original intensity and the path length of the light. Beer's law proposed that the transmittance of a solution remains constant if the product of concentration and path length is constant. The...
2.1K
Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview01:02

Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview

2.5K
Ultraviolet–visible (UV–visible or UV–Vis) spectroscopy is an analytical technique that investigates the interaction between matter and UV–Vis light within the electromagnetic spectrum. This method is widely used for its versatility, simplicity, and relatively quick data acquisition, making it valuable for both qualitative and quantitative analysis. When UV–Vis radiation passes through a material,  molecules absorb light depending on the energy required for...
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Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

537
Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
537

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Detection of Bacteria Using Fluorogenic DNAzymes
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Shining a Light on Spectrophotometry in Bacteriology.

Veronica Montesinos-Cruz1, Greg A Somerville1

  • 1School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0905, USA.

Antibiotics (Basel, Switzerland)
|January 8, 2025
PubMed
Summary

Spectrophotometry is a key tool for measuring bacterial growth. This review highlights essential principles and practical tips to ensure accurate and reproducible spectrophotometric data in microbiology experiments.

Keywords:
antibiotic resistancebacterial quantitationlight scatteringpersistencespectrophotometry

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Area of Science:

  • Microbiology
  • Biotechnology

Background:

  • Spectrophotometry is a common method for assessing bacterial growth in cultures.
  • Over-reliance on this technique can lead to complacency and experimental errors.

Purpose of the Study:

  • To review fundamental spectrophotometry principles for biological applications.
  • To provide practical considerations for improving the accuracy of growth measurements.
  • To help scientists generate robust and reproducible spectrophotometric data.

Main Methods:

  • Review of established spectrophotometry principles.
  • Discussion of practical experimental considerations.
  • Guidance on data interpretation and presentation.

Main Results:

  • Familiarity can breed complacency, impacting data reliability.
  • Several practical factors can influence spectrophotometric readings.
  • Adherence to best practices enhances data quality.

Conclusions:

  • Revisiting spectrophotometry fundamentals is crucial for accurate bacterial growth assessment.
  • Attention to practical details ensures robust and reproducible experimental outcomes.
  • This review serves as a guide for scientists using spectrophotometry in microbiology.