Inexpensive manual microscopy stages

I’m building a simple, low resolution microscope for imaging microfluidic devices (follow on research to our paper on bead encoding) and I wanted a cheap manual stage for the microscope.  All the stages I could find from the big microscopes vendors and the other usual suppliers of such things were over $1000, which is probably about what I spent on the rest of the components of the microscope. I wasn’t too happy about that, so I went looking for some cheaper options, and found a company, Manual Positioning, which makes a number of very cheap (<$200) microscope stages. One of their stages is resold by AmScope on Amazon with free prime shipping, which brings the cost down to only $80. I bought one, and it seems perfectly good for what I want to do. The control knob on the top of the stage is an odd choice, but not a problem for our application.

Stage-1

The stage, installed on our microscope.

To get it to work as I wanted I had to make some modifications – I milled out the mounting holes to use 8-32 screws for compatibility with Thorlabs posts, and replaced the glass plate that held the sample with a 3D-printed insert, but with that done it works very well.

Paper Roundup – December 2014

  • Using fluorogen-activating proteins (FAPs) to provide tunable labeling densities for ensemble imaging and single particle tracking [1]
  • Axial plane optical microscopy: using a remote objective with a tilted mirror to acquire an image of a tilted slice through your sample [2]
  • Measurement of axial scaling in confocal microscopy when imaging into samples of mismatched refractive index and comparison with theory [3]
  • Fast, multi-angle, circular scanning TIRF for getting Z-height information from TIRF imaging [4]
  • A photoswitchable long Stokes shift fluorescent protein [5]
  • A nice paper using single molecule imaging to look at enhancer binding to DNA in vivo [6]
  • A novel technique for measuring force by unwinding of a DNA hairpin containing a fluorescent dye and a quencher so that force causes the DNA molecule to light up [7]
  • A nice review of clearing techniques and objectives optimized for cleared samples [8]
  • Two super-resolution imaging systems operating over a 3-4μm range using a multiplane detection system [9] [10]
  • A paper studying the lethality of different wavelengths of light to different insects. They find that the most toxic wavelength is not always the shortest and that it varies from species to species. Possibly of interest to those thinking about phototoxicity. [11]
  • Correlative cryo-fluorescence microscopy and cryo-soft x-ray tomography [12]
  • A fluorescence turn-on approach for following RNAs by expression of an RNA aptamer and a cell-permeable fluorescent probe [13]
  • A pretty sophisticated approach to image transcription factor diffusion and binding in live cells [14]

References

  1. S.L. Schwartz, Q. Yan, C.A. Telmer, K.A. Lidke, M.P. Bruchez, and D.S. Lidke, "Fluorogen-Activating Proteins Provide Tunable Labeling Densities for Tracking FcεRI Independent of IgE", ACS Chemical Biology, vol. 10, pp. 539-546, 2015. http://dx.doi.org/10.1021/cb5005146
  2. T. Li, S. Ota, J. Kim, Z.J. Wong, Y. Wang, X. Yin, and X. Zhang, "Axial Plane Optical Microscopy", Scientific Reports, vol. 4, 2014. http://dx.doi.org/10.1038/srep07253
  3. T. BESSELING, J. JOSE, and A.V. BLAADEREN, "Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index", Journal of Microscopy, vol. 257, pp. 142-150, 2014. http://dx.doi.org/10.1111/jmi.12194
  4. J. Boulanger, C. Gueudry, D. Münch, B. Cinquin, P. Paul-Gilloteaux, S. Bardin, C. Guérin, F. Senger, L. Blanchoin, and J. Salamero, "Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging", Proceedings of the National Academy of Sciences, vol. 111, pp. 17164-17169, 2014. http://dx.doi.org/10.1073/pnas.1414106111
  5. K. Piatkevich, B. English, V. Malashkevich, H. Xiao, S. Almo, R. Singer, and V. Verkhusha, "Photoswitchable Red Fluorescent Protein with a Large Stokes Shift", Chemistry & Biology, vol. 21, pp. 1402-1414, 2014. http://dx.doi.org/10.1016/j.chembiol.2014.08.010
  6. J. Chen, Z. Zhang, L. Li, B. Chen, A. Revyakin, B. Hajj, W. Legant, M. Dahan, T. Lionnet, E. Betzig, R. Tjian, and Z. Liu, "Single-Molecule Dynamics of Enhanceosome Assembly in Embryonic Stem Cells", Cell, vol. 156, pp. 1274-1285, 2014. http://dx.doi.org/10.1016/j.cell.2014.01.062
  7. B.L. Blakely, C.E. Dumelin, B. Trappmann, L.M. McGregor, C.K. Choi, P.C. Anthony, V.K. Duesterberg, B.M. Baker, S.M. Block, D.R. Liu, and C.S. Chen, "A DNA-based molecular probe for optically reporting cellular traction forces", Nature Methods, vol. 11, pp. 1229-1232, 2014. http://dx.doi.org/10.1038/nmeth.3145
  8. V. Marx, "Microscopy: seeing through tissue", Nature Methods, vol. 11, pp. 1209-1214, 2014. http://dx.doi.org/10.1038/nmeth.3181
  9. B. Hajj, J. Wisniewski, M. El Beheiry, J. Chen, A. Revyakin, C. Wu, and M. Dahan, "Whole-cell, multicolor superresolution imaging using volumetric multifocus microscopy", Proceedings of the National Academy of Sciences, vol. 111, pp. 17480-17485, 2014. http://dx.doi.org/10.1073/pnas.1412396111
  10. S. Geissbuehler, A. Sharipov, A. Godinat, N.L. Bocchio, P.A. Sandoz, A. Huss, N.A. Jensen, S. Jakobs, J. Enderlein, F. Gisou van der Goot, E.A. Dubikovskaya, T. Lasser, and M. Leutenegger, "Live-cell multiplane three-dimensional super-resolution optical fluctuation imaging", Nature Communications, vol. 5, pp. 5830, 2014. http://dx.doi.org/10.1038/ncomms6830
  11. M. Hori, K. Shibuya, M. Sato, and Y. Saito, "Lethal effects of short-wavelength visible light on insects", Scientific Reports, vol. 4, 2014. http://dx.doi.org/10.1038/srep07383
  12. E. Smith, G. McDermott, M. Do, K. Leung, B. Panning, M. Le Gros, and C. Larabell, "Quantitatively Imaging Chromosomes by Correlated Cryo-Fluorescence and Soft X-Ray Tomographies", Biophysical Journal, vol. 107, pp. 1988-1996, 2014. http://dx.doi.org/10.1016/j.bpj.2014.09.011
  13. S. Sato, M. Watanabe, Y. Katsuda, A. Murata, D.O. Wang, and M. Uesugi, "Live-Cell Imaging of Endogenous mRNAs with a Small Molecule", Angewandte Chemie International Edition, vol. 54, pp. 1855-1858, 2014. http://dx.doi.org/10.1002/anie.201410339
  14. Z. Liu, W.R. Legant, B. Chen, L. Li, J.B. Grimm, L.D. Lavis, E. Betzig, and R. Tjian, "3D imaging of Sox2 enhancer clusters in embryonic stem cells", eLife, vol. 3, 2014. http://dx.doi.org/10.7554/eLife.04236