OSA's Digital Library

Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 4 — Apr. 1, 2012
  • pp: 735–740
« Show journal navigation

Three-dimensional, non-invasive, cross-sectional imaging of protein crystals using ultrahigh resolution optical coherence tomography

Norihiko Nishizawa, Shutaro Ishida, Mika Hirose, Shigeru Sugiyama, Tsuyoshi Inoue, Yusuke Mori, Kazuyoshi Itoh, and Hiroyoshi Matsumura  »View Author Affiliations


Biomedical Optics Express, Vol. 3, Issue 4, pp. 735-740 (2012)
http://dx.doi.org/10.1364/BOE.3.000735


View Full Text Article

Acrobat PDF (1144 KB)





Browse Journals / Lookup Meetings

Browse by Journal and Year


   


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools

Share
Citations

Abstract

Micro-scale, non-invasive, three-dimensional cross-sectional imaging of protein crystals was successfully accomplished using ultra-high resolution optical coherence tomography (UHR-OCT) with low noise, Gaussian like supercontinuum. This technique facilitated visualization of protein crystals even those in medium that also contained substantial amounts of precipitates. We found the enhancement of the scattered signal from protein crystal by inclusion of agarose gel in the crystallization medium. Crystals of a protein and a salt in the same sample when visualized by UHR-OCT showed distinct physical characteristics, suggesting that protein and salt crystals may, in general, be distinguishable by UHR-OCT. UHR-OCT is a nondestructive and rapid method, which should therefore find use in automated systems designed to visualize crystals.

© 2012 OSA

1. Introduction

X-ray crystallography is used to determine three-dimensional structures of proteins at atomic resolution. Protein crystals are required for this method, and automated robotic systems are currently used to prepare many different test media so that crystallization conditions can be rapidly surveyed. However, three dimensional imaging, which is important for automated treatment of protein crystal, is generally difficult by light microscopy. In addition, the crystals in these media are frequently difficult to identify by conventional light microscopy because the media often also contain salt crystals, e.g., NaCl, that appear similar to protein crystals, and/or protein crystals that are not suitable for X-ray crystallography, and/or amorphous materials that obscure the presence of usable protein crystals. Consequently, ultraviolet microscopy has been used to identify protein crystals [1

1. R. A. Judge, K. Swift, and C. González, “An ultraviolet fluorescence-based method for identifying and distinguishing protein crystals,” Acta Crystallogr. D Biol. Crystallogr. 61(1), 60–66 (2005). [CrossRef] [PubMed]

]; however, ultraviolet light can damage proteins [2

2. J. J. Kehoe, G. E. Remondetto, M. Subirade, E. R. Morris, and A. Brodkorb, “Tryptophan-mediated denaturation of beta-lactoglobulin A by UV irradiation,” J. Agric. Food Chem. 56(12), 4720–4725 (2008). [CrossRef] [PubMed]

].

Optical coherence tomography (OCT) is an emerging technique for μm-scale cross-sectional imaging [3

3. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991). [CrossRef] [PubMed]

5

5. W. Drexler and J. G. Fujimoto, Optical Coherence Tomography (Springer, 2008).

]. Generally, the axial resolution obtainable by OCT is ~10 μm and is dependent on the center wavelength and bandwidth of the light source. OCT is a non-destructive, rapid, three-dimensional imaging tool that has been used to visualize biological tissues and other materials. OCT has received much attention especially for medical imaging, and it has been used in ophthalmology clinics, etc. [3

3. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991). [CrossRef] [PubMed]

5

5. W. Drexler and J. G. Fujimoto, Optical Coherence Tomography (Springer, 2008).

]. The application for industrial field, such as the investigation of polymer matrix composites, has also been demonstrated [4

4. B. E. Bouma and G. J. Tearney, Handbook of Optical Coherence Tomography (Marcel Dekker, 2002)

,6

6. J. P. Dunkers, R. S. Parnas, C. G. Zimba, R. S. Peterson, K. M. Flynn, J. G. Fujimoto, and B. E. Bouma, “Optical coherence tomography of glass reinforced polymer composites,” Compos., Part A Appl. Sci. Manuf. 30, 139–145 (1999). [CrossRef]

]. The OCT signal is a beam backscattered from the sample, which makes the visualization of nearly transparent and low-scattering samples difficult.

Agarose gels have been included in protein crystallization media as they reduce solvent convection and prevent crystal sedimentation, which allows protein crystals to grow as if under microgravity [7

7. C. Biertümpfel, J. Basquin, D. Suck, and C. Sauter, “Crystallization of biological macromolecules using agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1657–1659 (2002). [CrossRef] [PubMed]

,8

8. B. Lorber, C. Sauter, M. C. Robert, B. Capelle, and R. Giegé, “Crystallization within agarose gel in microgravity improves the quality of thaumatin crystals,” Acta Crystallogr. D Biol. Crystallogr. 55(9), 1491–1494 (1999). [CrossRef] [PubMed]

]. When protein crystals are grown in agarose gels, they become trapped in the pores of the gel matrix. By immobilizing the crystals in a gel, the crystals can be subjected to processed by a femtosecond laser irradiation to mount onto the X-ray diffraction equipment for data collection [9

9. S. Sugiyama, H. Hasenaka, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of Agarose gel surrounding protein crystals for development of an automated Crystal capturing system,” Jpn. J. Appl. Phys. 48(10), 105502 (2009). [CrossRef]

,10

10. H. Hasenaka, S. Sugiyama, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of protein crystals grown in agarose gel,” J. Cryst. Growth 312(1), 73–78 (2009). [CrossRef]

]. Such crystals are more resistant to environmental perturbations, e.g., evaporation [11

11. S. Sugiyama, M. Hirose, N. Shimizu, M. Niiyama, M. Maruyama, G. Sazaki, R. Murai, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Effect of evaporation on protein crystals grown in semi-solid agarose hydrogel,” Jpn. J. Appl. Phys. 50(2), 025502 (2011). [CrossRef]

] and osmotic shock [12

12. C. Sauter, B. Lorber, and R. Giegé, “Towards atomic resolution with crystals grown in gel: the case of thaumatin seen at room temperature,” Proteins 48(2), 146–150 (2002). [CrossRef] [PubMed]

]. Furthermore, agarose gels promote protein nucleation [13

13. K. Tanabe, M. Hirose, R. Murai, S. Sugiyama, N. Shimizu, M. Maruyama, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, E. Mizohata, T. Inoue, and H. Matsumura, “Promotion of crystal nucleation of protein by semi-solid Agarose gel,” Appl. Phys. Express 2(12), 125501 (2009). [CrossRef]

,14

14. K. J. Thiessen, “The use of two novel methods to grow protein crystals by microdialysis and vapor diffusion in an agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 50(4), 491–495 (1994). [CrossRef] [PubMed]

].

Herein we report the first micro-scale, non-invasive, three-dimensional cross-sectional images of protein crystals by ultra-high resolution (UHR) OCT [15

15. W. Drexler, U. Morgner, R. K. Ghanta, F. X. Kärtner, J. S. Schuman, and J. G. Fujimoto, “Ultrahigh-resolution ophthalmic optical coherence tomography,” Nat. Med. 7(4), 502–507 (2001). [CrossRef] [PubMed]

,16

16. J. G. Fujimoto, A. D. Aguirre, Y. Chen, P. R. Herz, P.-L. Hsiung, T. H. Ko, N. Nishizawa, and F. X. Kartner, Ultrashort Laser Pulses in Biology and Medicine (Springer, 2007), Chap. 1.

]. In the developed UHR-OCT, the axial resolution of 2 μm in sample was achieved using a broadband supercontinuum (SC) source and specially optimized OCT at 800 nm wavelength region [17

17. M. Nishiura, T. Kobayashi, M. Adachi, J. Nakanishi, T. Ueno, Y. Ito, and N. Nishizawa, “In vivo ultrahigh-resolution ophthalmic optical coherence tomography using Gaussian-shaped super continuum,” Jpn. J. Appl. Phys. 49(1), 012701 (2010). [CrossRef]

,18

18. S. Ishida and N. Nishizawa, “Quantitative comparison of contrast and imaging depth of ultrahigh-resolution optical coherence tomography images in 800-1700 nm wavelength region,” Biomed. Opt. Express 3(2), 282–294 (2012). [CrossRef] [PubMed]

]. As mentioned above, OCT cannot detect transparent objects, e.g., ideal crystals, because such crystals have homogeneous nano-structures for which the scattering coefficient is small. As we report below, as expected, the OCT signal intensities of protein crystals that had been grown in solution were extremely small. Additionally, we found that the backscattered signals of protein crystals were enhanced when the crystals were grown in agarose gel-containing medium. Even when the crystallization medium contained substantial amounts of precipitants, such that the protein crystals could not been clearly seen by light microscopy, they were easily detectable by UHR-OCT. Finally, we found that the UHR-OCT images of salt and protein crystals differed, suggesting that the two types of crystals may routinely be differentiated by UHR-OCT

2. Methods and Materials

The interference signal was transferred to a personal computer to construct cross-sectional images. The cross-sectional image consists of 250 transverse scans with 1000 pixels per scan, covering an area of 2.0 mm by 1.0 mm. The imaging speed was 1 frame per second. The lateral resolution was 17.6 μm. For the imaging of protein crystals, the visible aiming beam was combined with SC and used to know the beam irradiation point. A CCD camera with zooming lens was also used to monitor the observation point on the sample (Fig. 1(c)).

Figure 1(b) shows a picture of protein crystals in a sample plate. For the preparation of protein crystals, Hen egg white lysozyme (HEWL, Seikagaku Corporation), agarose IX-A (Sigma-Aldrich), and 96-well microbatch plates (Hampton Research) were mainly used for crystallization [10

10. H. Hasenaka, S. Sugiyama, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of protein crystals grown in agarose gel,” J. Cryst. Growth 312(1), 73–78 (2009). [CrossRef]

]. HEWL (50 mg/ml) was crystallized by the batch method at 293 K in 0.33 M sodium acetate, 0.033 M sodium acetate (pH 4.5), 0.51 M sodium chloride, and agarose at 0.4% (w/v) increments between 0% and 2.0% (w/v) [13

13. K. Tanabe, M. Hirose, R. Murai, S. Sugiyama, N. Shimizu, M. Maruyama, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, E. Mizohata, T. Inoue, and H. Matsumura, “Promotion of crystal nucleation of protein by semi-solid Agarose gel,” Appl. Phys. Express 2(12), 125501 (2009). [CrossRef]

]. To prepare HEWL crystals in the substantial amounts of precipitants, HEWL (10 mg/ml) was crystallized in 0.25 M calcium chloride, 0.1 M citrate acetate, 0.066 M sodium acetate (pH 4.5), 0.85 M sodium chloride, 1.6% (w/v) agarose. To obtain salt and protein crystals simultaneously, the HEWL (12 mg/ml) was crystallized in 0.05 M calcium chloride, 0.1 M potassium phosphate, 0.066 M sodium acetate (pH 4.5), 0.85 M sodium chloride, 2.0% (w/v) agarose.

For comparison, Synechococcus phosphoribulokinase was purified as described [19

19. D. Kobayashi, M. Tamoi, T. Iwaki, S. Shigeoka, and A. Wadano, “Molecular characterization and redox regulation of phosphoribulokinase from the cyanobacterium Synechococcus sp. PCC 7942,” Plant Cell Physiol. 44(3), 269–276 (2003). [CrossRef] [PubMed]

] and crystallized by the sitting-drop method in 0.1 M 2-N-morpholino-ethanesulfonic acid buffer (pH 6.5), 10% (w/v) isopropanol, 0.2 M potassium acetate, 1% (w/v) agarose at 293 K.

3. Results and Discussion

We first used UHR-OCT to visualize the HEWL protein crystals grown in the presence of the different agarose concentrations to ascertain how the agarose gel affected the UHR-OCT images (Fig. 2
Fig. 2 Cross sections of three-dimensional UHR-OCT images of HEWL crystals grown in (a) 0.0, (b) 0.4, (c) 0.8, (d) 1.2, and (e) 1.8% (w/v) agarose. (f) An image of a drop that contained Synechococcus phosphoribulokinase crystals and 1.0% (w/v) agarose. The crystals shown in panels (e) and (f) are clearly and non-invasively seen at μm resolution. Media 1 shows the 3D UHR-OCT image of HEWL crystals grown in the same condition as that for Fig. 2(e).
). The positions of HEWL crystals and aiming beam were observed by CCD camera as shown in Fig. 1(c). When agarose was absent, crystals were hardly visible, probably because of their small backscattered signals (Fig. 2(a)). The backscattered signals were enhanced as the agarose concentration was increased (Fig. 2(a-e)), so that the crystals grown in the presence of 1.8% (w/v) agarose were clearly visible. The backscattered signals associated with the edges of the crystals were greatly enhanced (10-28 dB above the noise floor for the 1.8% agarose sample), which resulted in clear crystal profiles (Fig. 2(e)). The presence of the agarose gel therefore effectively enhanced the backscattered signals of the protein crystals. The average values for the backscattered signals for gel-grown HEWL crystals, agarose-gel-containing medium around the HEWL crystals, and solution-grown HEWL crystals were ~5, 2, and 0 dB above the noise floor, respectively. The standard deviation was 3 dB for a thousand sampling points in each part. We examined three sample plates of HEWL crystals and they showed almost the same results.

We also used crystallized Synechococcus phosphoribulokinase as a test case [19

19. D. Kobayashi, M. Tamoi, T. Iwaki, S. Shigeoka, and A. Wadano, “Molecular characterization and redox regulation of phosphoribulokinase from the cyanobacterium Synechococcus sp. PCC 7942,” Plant Cell Physiol. 44(3), 269–276 (2003). [CrossRef] [PubMed]

,20

20. A. Wadano, Y. Kamata, T. Iwaki, K. Nishikawa, and T. Hirahashi, “Purification and characterization of phosphoribulokinase from the cyanobacterium Synechococcus PCC7942,” Plant Cell Physiol. 36(7), 1381–1385 (1995). [PubMed]

]. The crystals were clearly visualized by UHR-OCT (Fig. 2(f)). Therefore, in general, agarose gels may enhance the backscattered signal, which would facilitate protein crystal visualization. Probably, the signal enhancement is a consequence of the different protein and agarose medium refractive indexes. As mentioned above, OCT signals are back-scattered at the boundaries of materials with distinct refractive indexes [4

4. B. E. Bouma and G. J. Tearney, Handbook of Optical Coherence Tomography (Marcel Dekker, 2002)

,5

5. W. Drexler and J. G. Fujimoto, Optical Coherence Tomography (Springer, 2008).

]. Agarose fibers penetrate protein crystals [21

21. J. A. Gavira and J. M. García-Ruiz, “Agarose as crystallisation media for proteins II: trapping of gel fibres into the crystals,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1653–1656 (2002). [CrossRef] [PubMed]

,22

22. S. Sugiyama, K. Tanabe, M. Hirose, T. Kitatani, H. Hasenaka, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, T. Inoue, and H. Matsumura, “Protein crystallization in Agarose gel with high strength: developing an automated system for protein crystallographic processes,” Jpn. J. Appl. Phys. 48(7), 075502 (2009). [CrossRef]

], suggesting that many boundaries between protein molecules and agarose fibers exist within the crystals, which would increase the intensity of the OCT signals.

According to the Rayleigh-scattering theorem, the scattering coefficient is proportional to d6/λ4, where d is the diameter of the scattering particles and λ is the wavelength of the irradiation beam. When protein crystals are grown in an agarose gel, the protein molecules are captured within the agarose-gel pores, which are ~300 nm in diameter [23

23. N. Pernodet, M. Maaloum, and B. Tinland, “Pore size of agarose gels by atomic force microscopy,” Electrophoresis 18(1), 55–58 (1997). [CrossRef] [PubMed]

]. Consequently, the protein molecules behave as small particles with diameters of a few hundred nm, which enhance the Rayleigh scattering signal.

Clinically, OCT is currently used to image semitransparent organs, e.g., the human eye, and practically opaque organs, e.g., human skin, internal organs, and blood vessels [3

3. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991). [CrossRef] [PubMed]

], which suggested to us that UHR-OCT could be used to visualize proteins that had been grown in agarose-gel-containing medium that also contained substantial amounts of opaque precipitants. Notably, visualization of crystals by light microscopy is difficult when the medium also contains a large amount of precipitant. To test our hypothesis, HEWL in the substantial amounts of precipitants was used as the sample. Indeed, identification of the HEWL crystals by light microscopy was difficult (Fig. 3(a)
Fig. 3 (a) Light micrograph. The HEWL crystal, circled in red, is difficult to see because it is surrounded by aggregates and amorphous material. (b) Cross section of a three-dimensional UHR-OCT image. The protein crystal is circled in red. The aggregates and amorphous material are colored black. The cross section is a still from Media 2.
), whereas the well-shaped HEWL crystals were clearly detected by UHR-OCT (Fig. 3(b)). UHR-OCT may therefore be a powerful tool for detecting protein crystals in medium containing precipitants.

4. Conclusion

In conclusion, micro-scale, non-invasive, three-dimensional cross-sectional imaging of protein crystals was demonstrated for the first time using ultra-high resolution optical coherence tomography (UHR-OCT) with Gaussian supercontinuum. The protein crystals grown in the presence of the different agarose gel concentrations were examined, and the enhancement of the scattered signal from protein crystal by inclusion of agarose gel was confirmed. Using the UHR-OCT and agarose gel inclusion technique, the protein crystals could be visible, even when the crystallization medium contains substantial amounts of precipitants. The signals intensities from a protein and a salt crystals were obviously different, suggesting that protein and salt crystals may be distinguishable. UHR-OCT is a nondestructive and rapid method (several sec/sample in principle), which can be incorporated into a robotic system that can rapidly screen for protein crystals suitable for X-ray crystallography.

References and links

1.

R. A. Judge, K. Swift, and C. González, “An ultraviolet fluorescence-based method for identifying and distinguishing protein crystals,” Acta Crystallogr. D Biol. Crystallogr. 61(1), 60–66 (2005). [CrossRef] [PubMed]

2.

J. J. Kehoe, G. E. Remondetto, M. Subirade, E. R. Morris, and A. Brodkorb, “Tryptophan-mediated denaturation of beta-lactoglobulin A by UV irradiation,” J. Agric. Food Chem. 56(12), 4720–4725 (2008). [CrossRef] [PubMed]

3.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991). [CrossRef] [PubMed]

4.

B. E. Bouma and G. J. Tearney, Handbook of Optical Coherence Tomography (Marcel Dekker, 2002)

5.

W. Drexler and J. G. Fujimoto, Optical Coherence Tomography (Springer, 2008).

6.

J. P. Dunkers, R. S. Parnas, C. G. Zimba, R. S. Peterson, K. M. Flynn, J. G. Fujimoto, and B. E. Bouma, “Optical coherence tomography of glass reinforced polymer composites,” Compos., Part A Appl. Sci. Manuf. 30, 139–145 (1999). [CrossRef]

7.

C. Biertümpfel, J. Basquin, D. Suck, and C. Sauter, “Crystallization of biological macromolecules using agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1657–1659 (2002). [CrossRef] [PubMed]

8.

B. Lorber, C. Sauter, M. C. Robert, B. Capelle, and R. Giegé, “Crystallization within agarose gel in microgravity improves the quality of thaumatin crystals,” Acta Crystallogr. D Biol. Crystallogr. 55(9), 1491–1494 (1999). [CrossRef] [PubMed]

9.

S. Sugiyama, H. Hasenaka, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of Agarose gel surrounding protein crystals for development of an automated Crystal capturing system,” Jpn. J. Appl. Phys. 48(10), 105502 (2009). [CrossRef]

10.

H. Hasenaka, S. Sugiyama, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of protein crystals grown in agarose gel,” J. Cryst. Growth 312(1), 73–78 (2009). [CrossRef]

11.

S. Sugiyama, M. Hirose, N. Shimizu, M. Niiyama, M. Maruyama, G. Sazaki, R. Murai, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Effect of evaporation on protein crystals grown in semi-solid agarose hydrogel,” Jpn. J. Appl. Phys. 50(2), 025502 (2011). [CrossRef]

12.

C. Sauter, B. Lorber, and R. Giegé, “Towards atomic resolution with crystals grown in gel: the case of thaumatin seen at room temperature,” Proteins 48(2), 146–150 (2002). [CrossRef] [PubMed]

13.

K. Tanabe, M. Hirose, R. Murai, S. Sugiyama, N. Shimizu, M. Maruyama, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, E. Mizohata, T. Inoue, and H. Matsumura, “Promotion of crystal nucleation of protein by semi-solid Agarose gel,” Appl. Phys. Express 2(12), 125501 (2009). [CrossRef]

14.

K. J. Thiessen, “The use of two novel methods to grow protein crystals by microdialysis and vapor diffusion in an agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 50(4), 491–495 (1994). [CrossRef] [PubMed]

15.

W. Drexler, U. Morgner, R. K. Ghanta, F. X. Kärtner, J. S. Schuman, and J. G. Fujimoto, “Ultrahigh-resolution ophthalmic optical coherence tomography,” Nat. Med. 7(4), 502–507 (2001). [CrossRef] [PubMed]

16.

J. G. Fujimoto, A. D. Aguirre, Y. Chen, P. R. Herz, P.-L. Hsiung, T. H. Ko, N. Nishizawa, and F. X. Kartner, Ultrashort Laser Pulses in Biology and Medicine (Springer, 2007), Chap. 1.

17.

M. Nishiura, T. Kobayashi, M. Adachi, J. Nakanishi, T. Ueno, Y. Ito, and N. Nishizawa, “In vivo ultrahigh-resolution ophthalmic optical coherence tomography using Gaussian-shaped super continuum,” Jpn. J. Appl. Phys. 49(1), 012701 (2010). [CrossRef]

18.

S. Ishida and N. Nishizawa, “Quantitative comparison of contrast and imaging depth of ultrahigh-resolution optical coherence tomography images in 800-1700 nm wavelength region,” Biomed. Opt. Express 3(2), 282–294 (2012). [CrossRef] [PubMed]

19.

D. Kobayashi, M. Tamoi, T. Iwaki, S. Shigeoka, and A. Wadano, “Molecular characterization and redox regulation of phosphoribulokinase from the cyanobacterium Synechococcus sp. PCC 7942,” Plant Cell Physiol. 44(3), 269–276 (2003). [CrossRef] [PubMed]

20.

A. Wadano, Y. Kamata, T. Iwaki, K. Nishikawa, and T. Hirahashi, “Purification and characterization of phosphoribulokinase from the cyanobacterium Synechococcus PCC7942,” Plant Cell Physiol. 36(7), 1381–1385 (1995). [PubMed]

21.

J. A. Gavira and J. M. García-Ruiz, “Agarose as crystallisation media for proteins II: trapping of gel fibres into the crystals,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1653–1656 (2002). [CrossRef] [PubMed]

22.

S. Sugiyama, K. Tanabe, M. Hirose, T. Kitatani, H. Hasenaka, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, T. Inoue, and H. Matsumura, “Protein crystallization in Agarose gel with high strength: developing an automated system for protein crystallographic processes,” Jpn. J. Appl. Phys. 48(7), 075502 (2009). [CrossRef]

23.

N. Pernodet, M. Maaloum, and B. Tinland, “Pore size of agarose gels by atomic force microscopy,” Electrophoresis 18(1), 55–58 (1997). [CrossRef] [PubMed]

OCIS Codes
(110.4500) Imaging systems : Optical coherence tomography
(170.3880) Medical optics and biotechnology : Medical and biological imaging

ToC Category:
Optical Coherence Tomography

History
Original Manuscript: February 7, 2012
Revised Manuscript: March 14, 2012
Manuscript Accepted: March 14, 2012
Published: March 15, 2012

Citation
Norihiko Nishizawa, Shutaro Ishida, Mika Hirose, Shigeru Sugiyama, Tsuyoshi Inoue, Yusuke Mori, Kazuyoshi Itoh, and Hiroyoshi Matsumura, "Three-dimensional, non-invasive, cross-sectional imaging of protein crystals using ultrahigh resolution optical coherence tomography," Biomed. Opt. Express 3, 735-740 (2012)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-3-4-735


Sort:  Author  |  Year  |  Journal  |  Reset  

References

  1. R. A. Judge, K. Swift, and C. González, “An ultraviolet fluorescence-based method for identifying and distinguishing protein crystals,” Acta Crystallogr. D Biol. Crystallogr. 61(1), 60–66 (2005). [CrossRef] [PubMed]
  2. J. J. Kehoe, G. E. Remondetto, M. Subirade, E. R. Morris, and A. Brodkorb, “Tryptophan-mediated denaturation of beta-lactoglobulin A by UV irradiation,” J. Agric. Food Chem. 56(12), 4720–4725 (2008). [CrossRef] [PubMed]
  3. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991). [CrossRef] [PubMed]
  4. B. E. Bouma and G. J. Tearney, Handbook of Optical Coherence Tomography (Marcel Dekker, 2002)
  5. W. Drexler and J. G. Fujimoto, Optical Coherence Tomography (Springer, 2008).
  6. J. P. Dunkers, R. S. Parnas, C. G. Zimba, R. S. Peterson, K. M. Flynn, J. G. Fujimoto, and B. E. Bouma, “Optical coherence tomography of glass reinforced polymer composites,” Compos., Part A Appl. Sci. Manuf. 30, 139–145 (1999). [CrossRef]
  7. C. Biertümpfel, J. Basquin, D. Suck, and C. Sauter, “Crystallization of biological macromolecules using agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1657–1659 (2002). [CrossRef] [PubMed]
  8. B. Lorber, C. Sauter, M. C. Robert, B. Capelle, and R. Giegé, “Crystallization within agarose gel in microgravity improves the quality of thaumatin crystals,” Acta Crystallogr. D Biol. Crystallogr. 55(9), 1491–1494 (1999). [CrossRef] [PubMed]
  9. S. Sugiyama, H. Hasenaka, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of Agarose gel surrounding protein crystals for development of an automated Crystal capturing system,” Jpn. J. Appl. Phys. 48(10), 105502 (2009). [CrossRef]
  10. H. Hasenaka, S. Sugiyama, M. Hirose, N. Shimizu, T. Kitatani, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Femtosecond laser processing of protein crystals grown in agarose gel,” J. Cryst. Growth 312(1), 73–78 (2009). [CrossRef]
  11. S. Sugiyama, M. Hirose, N. Shimizu, M. Niiyama, M. Maruyama, G. Sazaki, R. Murai, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, and H. Matsumura, “Effect of evaporation on protein crystals grown in semi-solid agarose hydrogel,” Jpn. J. Appl. Phys. 50(2), 025502 (2011). [CrossRef]
  12. C. Sauter, B. Lorber, and R. Giegé, “Towards atomic resolution with crystals grown in gel: the case of thaumatin seen at room temperature,” Proteins 48(2), 146–150 (2002). [CrossRef] [PubMed]
  13. K. Tanabe, M. Hirose, R. Murai, S. Sugiyama, N. Shimizu, M. Maruyama, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, E. Mizohata, T. Inoue, and H. Matsumura, “Promotion of crystal nucleation of protein by semi-solid Agarose gel,” Appl. Phys. Express 2(12), 125501 (2009). [CrossRef]
  14. K. J. Thiessen, “The use of two novel methods to grow protein crystals by microdialysis and vapor diffusion in an agarose gel,” Acta Crystallogr. D Biol. Crystallogr. 50(4), 491–495 (1994). [CrossRef] [PubMed]
  15. W. Drexler, U. Morgner, R. K. Ghanta, F. X. Kärtner, J. S. Schuman, and J. G. Fujimoto, “Ultrahigh-resolution ophthalmic optical coherence tomography,” Nat. Med. 7(4), 502–507 (2001). [CrossRef] [PubMed]
  16. J. G. Fujimoto, A. D. Aguirre, Y. Chen, P. R. Herz, P.-L. Hsiung, T. H. Ko, N. Nishizawa, and F. X. Kartner, Ultrashort Laser Pulses in Biology and Medicine (Springer, 2007), Chap. 1.
  17. M. Nishiura, T. Kobayashi, M. Adachi, J. Nakanishi, T. Ueno, Y. Ito, and N. Nishizawa, “In vivo ultrahigh-resolution ophthalmic optical coherence tomography using Gaussian-shaped super continuum,” Jpn. J. Appl. Phys. 49(1), 012701 (2010). [CrossRef]
  18. S. Ishida and N. Nishizawa, “Quantitative comparison of contrast and imaging depth of ultrahigh-resolution optical coherence tomography images in 800-1700 nm wavelength region,” Biomed. Opt. Express 3(2), 282–294 (2012). [CrossRef] [PubMed]
  19. D. Kobayashi, M. Tamoi, T. Iwaki, S. Shigeoka, and A. Wadano, “Molecular characterization and redox regulation of phosphoribulokinase from the cyanobacterium Synechococcus sp. PCC 7942,” Plant Cell Physiol. 44(3), 269–276 (2003). [CrossRef] [PubMed]
  20. A. Wadano, Y. Kamata, T. Iwaki, K. Nishikawa, and T. Hirahashi, “Purification and characterization of phosphoribulokinase from the cyanobacterium Synechococcus PCC7942,” Plant Cell Physiol. 36(7), 1381–1385 (1995). [PubMed]
  21. J. A. Gavira and J. M. García-Ruiz, “Agarose as crystallisation media for proteins II: trapping of gel fibres into the crystals,” Acta Crystallogr. D Biol. Crystallogr. 58(10), 1653–1656 (2002). [CrossRef] [PubMed]
  22. S. Sugiyama, K. Tanabe, M. Hirose, T. Kitatani, H. Hasenaka, Y. Takahashi, H. Adachi, K. Takano, S. Murakami, Y. Mori, T. Inoue, and H. Matsumura, “Protein crystallization in Agarose gel with high strength: developing an automated system for protein crystallographic processes,” Jpn. J. Appl. Phys. 48(7), 075502 (2009). [CrossRef]
  23. N. Pernodet, M. Maaloum, and B. Tinland, “Pore size of agarose gels by atomic force microscopy,” Electrophoresis 18(1), 55–58 (1997). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

Figures

Fig. 1 Fig. 2 Fig. 3
 
Fig. 4
 

Supplementary Material


» Media 1: MOV (3829 KB)     
» Media 2: MOV (3999 KB)     
» Media 3: MOV (3903 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited