METHOD AND APPARATUS FOR MEASURING BIOFILM THICKNESS AND TOPOLOGY
An apparatus and method of measuring biofilm and biological activity on a surface is disclosed. The apparatus includes a biofilm, which includes one or more microorganisms, grown on a substrate. A viewing window is placed on a surface of the biofilm and a gas bubble is introduced between the viewing window and the surface of the biofilm. The space between the substrate and the viewing window may be enclosed in a casing that has an inlet and an outlet, forming a flow cell. A microscope system, such as a white light interferometer, captures data of the biofilm in situ and non-destructively. The 3D images of biofilm surface have high resolution while maintaining a large field of view. This invention will be useful for fundamental studies of biofilms, biomedical and environmental screening, and many other applications in biology and the life sciences.
1 . An apparatus for measuring biofilm surface properties with a microscope system comprising:
a. a biofilm including one or more microorganisms grown on a substrate;
b. a viewing window placed on a surface of the biofilm; and
c. a gas bubble positioned between the viewing window and the surface of the biofilm for localizing a field of view.
2 . The apparatus of claim 1 wherein the substrate includes a reflective surface.
3 . The apparatus of claim 1 wherein the microscope system is a three-dimensional profile microscope system.
4 . The apparatus of claim 3 wherein the three-dimensional profile microscope system is a white light interferometer which generates and captures interference fringes of the biofilm.
5 . The apparatus of claim 1 wherein the biofilm, viewing window and gas bubble are located within a casing that has an inlet and an outlet and forms a flow cell.
6 . The apparatus of claim 5 wherein the flow cell is coupled to at least one pump that provides a gas bubble to the biofilm.
7 . The apparatus of claim 6 further comprising a second pump that provides ing the liquid media to a region between the substrate and the window, and a second pump for introducing the gas bubble to the biofilm.
8 . The apparatus of claim 7 wherein the apparatus comprising the flow cell includes a plurality of individual flow cells.
9 . An apparatus for measuring biofilm surface properties with a microscope system comprising:
a. a flow cell including an inlet, an outlet, a window, and a substrate;
b. a biofilm including one or more microorganisms grown on the substrate of the flow cell; and
c. a gas bubble positioned between the window and a surface of the biofilm.
10 . The apparatus of claim 9 wherein the flow cell is coupled to one or more pumps that provide a supply of liquid media and gas bubble to the biofilm.
11 . The apparatus of claim 10 wherein the one or more pumps includes a first pump for providing the liquid media to a region between the substrate and the window, and a second pump for introducing the gas bubble to the biofilm.
12 . The apparatus of claim 11 wherein the apparatus comprising the flow cell includes a plurality of individual flow cells.
13 . The apparatus of claim 12 wherein the microscope system is a three-dimensional profile microscope system.
14 . The apparatus of claim 13 wherein the three-dimensional profile microscope system is a white light interferometer.
15 . A method of measuring biofilm surface properties comprising:
a. growing one or more microorganisms on a substrate to form a biofilm;
b. placing a viewing window on a surface of the biofilm;
c. introducing a gas bubble between the viewing window and the surface of the biofilm; and
d. capturing data of the biofilm using a microscope system.
16 . The method of claim 15 further comprising monitoring changes in topology and thickness of the biofilm in response to changes in a local environment, wherein the local environment is the structure, composition, and chemistry of the viewing window and the substrate.
17 . An apparatus for measuring biofilm surface properties with a microscope system comprising:
a. a multi-well plate containing a plurality of individual wells, each well having a base;
b. a biofilm including one or more microorganisms grown on the base of the wells; and
c. a gas bubble localized on a surface of the biofilm.
18 . The apparatus of claim 17 wherein the well plate is made of a transparent material.
19 . The apparatus of claim 17 wherein the wells are positioned in an inverted orientation.
20 . The apparatus of claim 17 wherein the microscope system is a three-dimensional profile microscope system.
21 . The apparatus of claim 20 wherein the three-dimensional profile microscope system is a white light interferometer.