Methods

Name

Description

Target

Ref.

Qualitative and indirect

Biofilm Ring Test

Measuring the lack of microbead spots magnetic field and the mobility of the beads when combined with a bacterial solution in a polystyrene microplate indicate the presence of a biofilm.

The characterization of biofilm-forming bacteria

[83]

Steam-based method

To maintain biofilm integrity and increase the repeatability of biofilm measurement, steam is employed as a mild cleaning technique.

It enables the measurement of the live bacteria in the biofilm and the differentiation between strains that form biofilm and strains that don't develop biofilm.

[84]

Tube method (TM)

The detection of visible biofilm

Observation of biofilm lining the bottom and the walls of the tube for the qualitative detection

[85]

Congo red agar (CRA)

Colonies inoculated on CRA media change colour.

Qualitative detection by observing colony colour change

[86]

Roll plate method

Microorganisms colonize the external surface and visualize indirectly

Detection of colonization

[87]

Quantitative and   artificial techniques

AI-driven image analysis

The colour shift of injected colonies on CRA medium

Segmenting the bacterial cells and extracting the microscale geometric characteristics of the biofilm

[88]

A Multi-disciplinary Procedure

Multiple existing data sets from related studies were combined to analyze meta-data about biofilm development.

Understanding the formulation process can help to create successful biofilm control techniques.

[89]

Scanning electron microscopy images

Separating biofilm from scanning electron microscopy pictures using machine learning methods.

To detect the formation of biofilm and destruction of biofilm

[90]

Deep Learning Approach

Convolutional neural networks (CNNs) were utilized to accurately identify recent photographs using a small number of EFM images depicting biofilms of varying bacterial compositions. CNN performance was compared to that of human experts.

To detect Biofilms of different bacterial compositions

[91]

Epifluorescence microscopy (EFM) on mineral sulfides

Combination with bacterial species-specific nucleic acid dyes

Enables the detection of particular groups of microorganisms adhering to metal sulfide surfaces and allows for the investigation of biofilm structure and spatial distribution of cells.

[92]

Open-source robotic platform (EvoBot)

By carrying out completely automated biofilm cultivations with the possibility of using optical coherence tomography to see the biofilms (OCT) 

To detect reliable parameters describing biofilm structure.

[93]

Confocal microscopy images

Confocal laser scanning microscopy (CLSM) selectively stimulates fluorescence signals from different planes inside a sample and takes point-by-point photographs using localized laser excitation at specific wavelengths.

To detect the presence of extracellular DNA, exopolysaccharides, and biofilm viability

[93]

BiofilmQ

The technical details are presented in the online documentation based on conventional image analysis methods, novel object tracking, and image cytometry algorithms.

Numerous internal and external biofilm features may be quantified, analyzed, and visualized in three dimensions in time and space.

[94]

Time-lapse imaging method

Bright-field stereomicroscope with episcopic illumination and a microcontroller system that synchronizes a robotic arm with the movement of a stereomicroscope-mounted camera and lighting conditions are used in an Arduino-based method for time-lapse imaging of the whole biofilm ontogeny.

To detect biofilm development of different morphological and developmental dynamics

[95]

Microtiter plate (MP)

Biofilm detection with a microplate reader

Quantitative detection of biofilm

[85]

Real-time PCR

Pathogens can be identified by amplifying species-specific nucleic acid sequences, and virulence factors can be detected by amplifying target virulence genes, such as biofilm genes, using gene-specific primers.

Detection of biofilm genes

[85]

Conventional PCR

Multiplex PCR