ibidi : µ-Slide 4 Well Ph+ Glass Bottom

A chambered coverslip with 4 wells, a #1.5H glass bottom, and a special intermediate plate for excellent phase contrast, suitable for use in TIRF and single molecule applications

Perfect cell imaging thanks to the low thickness variability of the coverslips’ glass
Cost-effective experiments using small numbers of cells and low volumes of reagents
Excellent phase contrast over the entire well – no meniscus
Surface Modification: #1.5H (170 µm +/- 5 µm) D 263 M Schott glass, sterilized
Pcs./Box: 15 (individually packed)

Applications

Cultivation and microscopy of cell cultures
TIRF and single molecule applications

Specifications

Outer dimensions (w x l)	25.5 x 75.5 mm²
Number of wells	4
Dimensions of wells (w x l x h)	21.6 x 11.4 x 9.3 mm³
Volume per well	700 µl
Liquid height	3.0 mm
Total height with / without lid	10.8 / 9.5 mm
Growth area per well	2.5 cm²
Coating area per well	5.9 cm²
Bottom: Glass coverslip No. 1.5H, selected quality, 170 µm +/- 5 µm

Technical Features

Open µ-Slide with 4 independent wells
Intermediate plate in each well to avoid meniscus formation
Ph+ feature for perfect phase contrast—no meniscus formation
Bottom made from D 263 M Schott glass, No. 1.5H (170 +/- 5 µm)
May require coating to promote cell attachment
Also available as a µ-Slide 4 Well^Ph+ with an ibidi Polymer Coverslip Bottom for superior cell growth

Experimental Example

Surface-near F-actin network of a Dictyostelium discoideum DdLimE-GFP cell. Live cell imaging on a Glass Coverslip #1.5H using Total Interference Reflection Fluorescence (TIRF) microscopy.

ibidi Polymer Coverslip vs. ibidi Glass Bottom

	ibidi Polymer Coverslip	ibidi Glass Bottom
Optical properties
Refractive index (n_D 589 nm)	1.52	1.52
Abbe number	56	55
Thickness	#1.5 (180 µm)	#1.5H (170 µm)
Material	Microscopy plastic	D 263M Schott borosilicate glass
Autofluorescence	Low	Low
Transmission	Very high (even ultraviolet)	High (ultraviolet restrictions)
Birefringence (DIC)	Low (DIC compatible*)	Low (DIC compatible*)

Other aspects
Surface modifications	ibiTreat – tissue culture treated Uncoated – hydrophobic	Only glass
Protein coatings	Compatible	Compatible
Gas permeable	Yes	No
Material flexibility	High	Low
Breakable	No	Yes
Applications	Fluorescence microscopy	TIRF and single photon

* Special glass DIC Lids are available separately and open µ-Slides are available separately.

Phase Contrast Microscopy

Phase contrast microscopy is the most commonly used, transmitted light technique in cell culture. When working with phase contrast microscopy, it is crucial to have the two phase rings adjusted to each other. In open wells, the meniscus at the air-water-interphase works like a lens that refracts the beam path. This miscalibrates the phase rings, leading to poor contrast in the microscopic image.

Working with the µ-Slide 4 Well^Ph+ diminishes the meniscus, so that the whole optical system is aligned, no matter which position of the well is being imaged.

The µ-Slide 4 Well^Ph+ Glass Bottom (Phase Contrast +) is designed for excellent optical quality, especially for cell culture when normal phase contrast microscopy is being used. Opposed to the classic µ-Slide 4 Well Glass Bottom, the Ph+ version provides a special intermediate plate in the center of the well. This plate flattens the meniscus that disturbs the phase contrast effect in normal open wells. Openings near the corners provide access to the wells for easy filling and aspiration of liquids.

This innovative technique supports meniscus-free phase contrast microscopy in a very convenient manner.

Working with the µ-Slide 4 Well^Ph+ Glass Bottom diminishes the meniscus and increases the area of well contrasted cells

The illustration on the left shows the perturbing effect of a meniscus. Light is refracted on the air-water-interface, leading to poor contrast in microscopy. Only the small center part exhibits satisfying phase contrast.

Working with the µ-Slide 4 Well^Ph+ Glass Bottom diminishes the meniscus and increases the area of nicely contrasted cells. This nice contrast is due to the parallel beam path that is created by the plate.